Dual release nicotine formulations, and systems and methods for their use

ABSTRACT

This invention relates generally to a method to provide habitual tobacco users with products, methods and apparatus to reduce and eventually terminate their dependence on nicotine containing products. More specifically, the invention relates to a nicotine-based medicament that is formulated in such a way as to effectively reduce or eliminate the sensations of craving associated with addictive nicotine use.

PRIORITY DOCUMENTS

This application claims the benefit of U.S. Provisional Application Nos.60/982,070, filed Oct. 23, 2007; 60/868,238, filed Dec. 1, 2006;60/911,044, filed Apr. 10, 2007; 60/913,185, filed Apr. 20, 2007;60/916,510, filed May 7, 2007; and 60/917,190, filed May 10, 2007 and isa continuation-in-part of U.S. application Ser. No. 11/097,598 filed onApr. 1, 2005 which is a continuation of Ser. No. 10/913,103, filed Aug.6, 2004, issued Apr. 5, 2005, as U.S. Pat. No. 6,874,507, which is adivisional of Ser. No. 10/147,390, filed May 15, 2002, issued Oct. 5,2004, as U.S. Pat. No. 6,799,576, which is a continuation-in-part ofSer. No. 09/611,423, filed Jul. 7, 2000, now abandoned, which claimsbenefit to U.S. Provisional Application No. 60/144,140 filed on Jul. 16,1999, all of which applications are incorporated herein by reference.

FIELD OF THE INVENTION

This invention relates generally to compositions and methods allowinghabitual smokers to reduce and eventually terminate their dependence onnicotine containing products, particularly tobacco products. Morespecifically, the invention relates to a nicotine-based medicament thatis formulated in such a way as to effectively reduce or eliminate thesensations of craving associated with addictive smoking behavior.

BACKGROUND

The use of tobacco products presents a critical international publichealth problem. Addiction to nicotine represents an enormous health,social, and financial burden. Cigarettes are among the most addictiveproducts known, and a vast majority of people who attempt to quitsmoking relapse within days (Henningfield, 1991). They are the world'sleading cause of preventable death, contributing to 5 million prematuredeaths in 2000, and is estimated to increase to 10 million by 2020(Nides, 2006). In the United States, fewer than 10% of the nearly 20million people who quit smoking remain abstinent one year later. Thus,only 2-3% of smokers become non-smokers each year (Henningfield, 1995).

Nicotine is the primary active ingredient in cigarettes that reinforcesindividual smoking behavior. Nicotine interacts with nicotiniccholinergic receptors in the brain to induce the release ofneurotransmitters and produce an immediate reward—the “rush” thatsmokers experience—that is associated with a rapid rise in blood level.A persistent stimulus is also produced, and is associated with a highblood level of nicotine. As such, the dopaminergic reward system isactivated by nicotine which eventually results in nicotine dependency.However, it is the other constituents of tobacco and not nicotine thatcause widespread mortality and morbidity.

Nicotine replacement therapies (NRTs) are pharmacological nicotinedelivery systems developed to improve outcomes in tobacco cessationtreatment. Abrupt cessation of tobacco use often produces a withdrawalsyndrome that includes depression and/or anxiety, hunger, sleepdisruption, and inability to concentrate (Hughes, 1986). Withdrawalsymptoms usually peak within a few days of cessation and can last for upto 4 weeks. More than half the smokers who quit will relapse within oneweek, coinciding with the peak in withdrawal symptoms (Henningfield,1995).

NRTs help with smoking cessation by reducing the severity of withdrawalsymptoms, uncoupling the behavioral changes needed to quit from theunpleasant effects of nicotine withdrawal, thereby partially enhancingmood and improving concentration.

There are now a number of approved nicotine-containing smoking cessationproducts available by prescription starting with the launch of a gum in1984 and with OTC (over-the-counter) products available since about1996. Smoking cessation products in the U.S. are available in gums,patches, lozenges, and an inhaler and a nasal spray and use eithernicotine or nicotine derivatives.

Nicorette® gum (nicotine polacrilex) was approved for prescription salein 1984, and the FDA began allowing its sale without a prescription inFebruary 1996. The nicotine patch (containing nicotine free base), alsoknown as a nicotine transdermal system, has been available in the US byprescription since 1991 and by OTC since July 1996. It is sold under thebrand names Nicoderm®, Nicotrol®, Habitrol®, and Prostep®. The firstinhaled dosage form of nicotine, Nicotrol NS®, designed to be used as anasal inhaler, was launched in September 1996. FDA approved theNicotrol® nicotine inhalation system (containing nicotine free base) forsmoking cessation in May 1997. Nicotine enters the user's mouth througha mouthpiece attached to a plastic cartridge. Although termed an“inhaler,” it does not deliver nicotine to the lungs the way a cigarettedoes. Almost all of the nicotine travels only as far as the mouth andthroat, where it is absorbed through the mucous membranes (Schneider,2001). Nicotrol® nicotine nasal spray was approved in March 1996 forsale by prescription. The nicotine is inhaled into the nose from a pumpbottle and absorbed through the nasal lining into the bloodstream. Eachform of NRT has its own advantages and limitations.

Obtaining nicotine from NRT is considerably safer than doing so fromcigarettes, as the user is not exposed to any of the myriad harmfulcompounds of tobacco combustion. Although long-term use of NRT is notthought to be associated with any serious harmful effects (Molyneux,2004), the current forms of NRT smoking cessation products have verypoor efficacy; a recent survey has suggested that the percentage ofabstinent smokers varies between 14-24% for the various NRTs (Silagy,2004). No existing NRT offers the pharmacokinetic properties thatadequately satisfy the subjective craving that smokers experience andwhich would allow smokers to easily transition from a tobacco product toa safer NRT product from which he or she may begin to gradually reducehis or her dependence toward eventual cessation of all nicotine andtobacco containing products.

Low nicotine replacement levels or under-dosing and the inability toadequately satisfy a smoker's craving is likely a significant factorleading to the failure of many NRTs: “Dependence on smoking appears tobe related, at least in part, to the achievement of a rapid rise inplasma nicotine concentrations. If this assessment is correct, the mostdesirable adjuvant for smoking cessation would be one that closelymimics this pattern of plasma nicotine concentrations” (Svensson, 1987).Optimal nicotine replacement for smoking cessation may initially requireboth reproducing and then sustaining the nicotine levels in the bloodstream and in the brain that are produced by habitual cigarette smokingso that nicotine withdrawal symptoms are minimized while the behavioralaspects of smoking are modified towards cessation (Russell, 1986;Perkins, 1986). The most important factor in successful smokingcessation may be the ability to approximate the plasma nicotineconcentration pharmacokinetics obtained with cigarettes in order tosatisfy the subjective cravings. For example, in contrast to thecurrently available slow-release nicotine polacrilix gums (the nicotineis bound or complexed with the polacrilex resin), there is evidence thatrapid-release nicotine gum reduces craving more rapidly (see Niaura etal, Addiction 2005 100; 1720-1730).

It is also difficult to reproduce the pharmacokinetic pattern ofmultiple nicotine concentration peaks that are achieved by successive“puffs” from a cigarette, cigar, or pipe without the use of multipledoses. Thus, the slow rise and lack of achieving an adequate plasmanicotine concentration “steady state” around the time ofself-administration suggests a pharmacokinetic explanation for therelatively high failure rate of some NRTs; administering one large doseequivalent to the total nicotine inhaled from a cigarette or othertobacco containing product could be dangerous and it would only providea short duration of the high nicotine concentration in the blood stream.Providing smokers with nicotine in a form that does not requireinhalation of tobacco smoke could be an effective way to avoid thehazards associated with smoking.

There are as yet no currently approved products that are able to achievehigh peak nicotine concentrations combined with the benefits ofproviding a sustained, slow-release plasma level over a prolonged periodof time. U.S. Pat. No. 5,935,604 describes a nasal drug deliverycomposition comprising nicotine or a pharmacologically-acceptable saltor derivative wherein the composition is adapted to delivery of a pulseof nicotine for rapid absorption and a controlled release of nicotinefor subsequent sustained absorption. However, the small surface area ofthe nasal cavity does not afford the same opportunities as the largesurface area of the respiratory tract with surfaces and anatomicallocations varying in their absorption barrier properties.

There is a clear need for improvement in nicotine cessation treatment.Development of new systems is critical in an effort to both bypasslimitations of existing systems and to provide effective options formatching smokers to treatment.

SUMMARY

The present invention provides tobacco-less formulations and methodsallowing a person to overcome a smoking habit. The invention providesthe smoker with a combination dosage form that has readily bioavailablenicotine causing rapidly high “peak” nicotine levels that are thought tobe associated with “craving” for nicotine, and a slow release componentof the formulation that maintains the nicotine plasma levels over aprolonged period of time. The dual release nature of this formulation,the rapid release component to achieve a quick peak nicotine plasmaconcentration, and the slow-release component to achieve lower,sustained plasma concentrations, will allow the subjective cravingsexperienced by the smoker to be minimized. Such formulations could beadministered by injection, or inhalation into the airways and lungs viamouth or nose, or applied to the nasal mucosa or swallowed or giventransdermally (by a combination of e.g., microneedles plus a slowrelease patch), or via the buccal cavity, etc.

The rapid-release component could be nicotine, or a nicotine-likesubstance, or a salt of nicotine, in liquid form, or dissolved in asuitable solvent, or be in a rapidly soluble solid form. To achieverapid absorption, a penetration enhancer may be added. The slowreleasing component could be a liposomal formulation, or a cyclodextrincomplex, or nicotine in a solid matrix slowly releasing (e.g., apolylacticglycolic acid microsphere, an ion-exchange resin such aspolacrilex [Amberlite IRP64], or lipid-based microspheres). The mixtureof the slow releasing and the rapidly-releasing components (e.g., anicotine salt solution in water together with nicotine-loaded liposomesin the same aqueous formulation; or nicotine dissolved in a suitablepropellant plus nicotine-containing slow-release particles dispersed ina propellant, the mixture being enclosed in a “metered dose inhaler”)would be administered into the body, e.g., by inhalation of anaerosolized mixture. Methods of formulating liquids and liquid inhalersare disclosed in U.S. Pat. Nos. 5,364,838; 5,709,202; 5,497,763;5,544,646; 5,718,222; 5,660,166; 5,823,178; and 5,910,301; all of whichare incorporated by reference to describe and disclose such.Formulations for both rapid and slow release of nicotine include aqueousformulations, aqueous saline formulations, and ethanol formulations. Adry powder formulation comprising a pharmacologically acceptable salt ofnicotine alone or with additives such as components to prevent theparticles from sticking together may be used.

All of these formulations may be included with additional componentssuch as permeation enhancers, buffers, preservatives and excipient andcarrier components and additives normally included within formulationsfor aerosolized drug delivery.

The advantages of this nicotine product would be that a singleadministration would achieve a safe and effective peak of nicotine inthe blood stream and then maintain the concentration of nicotine at asuitable level for prolonged period of time to avoid “craving” fortobacco products. This combination product could be used both as asmoking cessation tool as well as a safer replacement for tobaccosmoking.

In addition, an aspect of the invention is that varying the amounts ofeach of the dual release formulation components, makes it possible togradually reduce the blood concentrations of nicotine. In this way, thesmoker is able to achieve eventual freedom from dependence on nicotineand any nicotine containing tobacco product.

In one embodiment the invention provides a tobacco-less composition thathas a pharmaceutically active nicotine formulation for delivery to apatient. The nicotine formulation has at least two forms of nicotine, afast release form and a slow release form. At least the fast releaseform of nicotine is inhaled, and is present in an amount to provide afirst form of nicotine arterial concentration in the patient within 5minutes of delivery. The nicotine arterial concentration produced by thefirst nicotine form is preferably at least 10 ng/ml, but may be 15, 20,25, 30, 35, 40, 50 or more ng/ml nicotine, being limited by the amountnecessary to address the nicotine addiction while not reaching toxiclevels.

The second form of nicotine is present in an amount to maintain a secondform of nicotine arterial concentration in the patient for at least 60minutes after delivery. This may be augmented by addition of slowrelease components such as cyclodextrin, encapsulation of the activenicotine, chemical or physical modification of the form of nicotine andthe like. The arterial concentration of the second form of nicotine isat least 5 ng/ml, preferably 7, 10, 12, 15 or 20 ng/ml.

Encapsulation of the second form of nicotine may be by any method knownin the art. For example, the nicotine form may be encapsulated in amicrosphere, such as a polyglycolide microsphere, or a liposome. Theform of nicotine may be further modified by optional addition of abioadhesive component, such as hyaluronic acid. Encapsulation greatlyenhances the variation in nicotine forms that may be utilized in thetobacco-less formulation. For example, by creating controlledmicroenvironments, encapsulation allows different pH forms, differentsalts and different compounds to be associated with one form of nicotinewithout contamination of the other. For example, by allowing eachnicotine form to be optionally encapsulated, the tobacco-lessformulation may include nicotine forms delivered at different pH values.This is important as the pH of the solution containing the compounddetermines whether the compound is a free base, acid or salt. It is wellknown in the art that free base nicotine is much more potent than saltsin eliciting a nicotine response in humans. Thus by delivering the firstform of nicotine at a basic pH and the second form of nicotine at anacidic pH augments the invention in providing a greater effectivenicotine in the initial nicotine bolus while also augmenting the slowrelease aspect of the second nicotine form.

In certain embodiments, the first and second forms of nicotine may alsobe packaged and/or delivered to the patient separately. Theseembodiments are distinct from those having the first and second forms ofnicotine present in the same tobacco-less formulation. Packaging the twoforms of nicotine separately allows the first form to be inhaled oradministered via another route affording an initial pulse of nicotine tothe arterial circulation, with the second form being delivered orally,transdermally or via some route consistent with the slow release natureof the second form. Lozenges, gums and quick dissolve strips are justsome examples of compositions suitable for oral administration, withmore examples presented below.

Pharmaceutically active nicotine formulations of the invention may becreams, gels, solids, patches, lozenges, gums, fast dissolving strips,powders, liquids, suspensions, emulsions and the like. In someembodiments the nicotine formulation is suitable for forming an aerosol.Such aerosols may include a propellant or be driven by mechanicalmanipulation without inclusion of a propellant. With aerosolizedembodiments of the claimed invention the first form of nicotinetypically has a smaller particle diameter than the second form ofnicotine. For example, the first form of nicotine may have a particlediameter between about 1 μm and about 4 μm, more preferably betweenabout 2 or 3 μm in diameter. Second form of nicotine particles typicallyhave a diameter between about 4 μm and about 12 μm

Tobacco-less formulations of the invention may also includeantidepressant or anxiolytic compounds or other supplemental drugs,excipients or other compounds that enhance the formulation chemically,pharmaceutically or in its ease or pleasure of use. In theory, theantidepressant or anxiolytic should allow the patient to make that finaltransition off nicotine entirely more easily.

The present invention also includes methods for treating a patient withtobacco-less nicotine formulation described above. The methods includedelivering the nicotine formulation to a patient.

One aspect of the invention is a method of treatment, comprising:

(a) aerosolizing a formulation comprised of nicotine creatingaerosolized particles which are sufficiently small to target and depositpredominantly in a particular lower area of the respiratory tract suchas the alveoli. The particles targeting this area will have a relativelysmall size, e.g. 0.5 micron to about 2 microns in diameter.

(b) in the next step the patient inhales the aerosolized particles of(a) into the respiratory tract, preferably targeted to a specific areaof the lower respiratory tract where the deposited particles cross intothe patient's circulatory system.

In step (c), steps (a) and (b) are repeated a plurality of times.Specifically, the patient may repeat these steps any number of timessuch as every time the patient would normally smoke a cigarette. At thispoint the patient could continue the treatment protocol in this mannerand gradually decrease the number of times the patient administersaerosolized nicotine until the patient is no longer addicted tonicotine. Decreasing the amount of aerosolized nicotine could also bedone by decreasing the concentration of nicotine within the aerosolizedparticles by decreasing the concentration of nicotine in the formulationand/or decreasing the size of the aerosolized dose.

Preferably the method of the invention continues with a step (d) whichinvolves aerosolizing formulation comprised of nicotine in order tocreate aerosolized particles which are larger in size than theaerosolized particles produced in step (a). These larger particles aredirected towards a particular area of the patient's respiratory tract,e.g. the mid-region of the patient's respiratory tract. (See FIGS. 1 and2) These particles could have a size in the range of about 2 microns toabout 4 microns.

In the following step (d) the patient inhales the aerosolized particlesof (d) thereby targeting the particular desired area of the patient'srespiratory tract such as the mid region. Thereafter, steps (d) and (e)are repeated a plurality of times. At this point the patient candecrease the amount of nicotine being delivered as indicated in the samemanner as indicated above step (c). Alternatively, the method of theinvention can be continued so that a third phase of treatment can becarried out which phase is similar to the two phases described above. Inaccordance with the above invention it is possible to carry out thetreatment in any number of phases. It would be impractical to develop asystem which attempts to target each of the 24 different areas of thelung as outlined in Table 1 and shown in FIG. 1. Further, regardless ofthe system used there would be some overlap between the different areasof the lung. Because it may not be practical to specifically design theparticles so that they are all larger in each of the phases theformulations may be designed so that a certain percentage of theparticles within each phase of delivery is larger than the particles inthe preceding phase.

Methods of the invention also include a method for treating a patientwith the pharmaceutically active, tobacco-less nicotine formulationdescribed. The method includes delivering the nicotine formulation to apatient in a first dosage amount and determining the patient's cravingfor nicotine after administering the formulation. The patient's cravingfor nicotine may be determined using any method known in the art,preferably the Fagerstrom test as described below. A second dosageamount of the formulation may be optionally administered to the patient.This second dosage amount may be a larger or smaller amount of thetherapeutically effective nicotine formulation than was administered inthe first dosage amount.

Another embodiment of the method of the invention involves treating apatient with a pharmaceutically active, tobacco-less nicotineformulation. The method includes delivering the nicotine formulationdiscussed herein to a patient over a first period of time in a firstdosage amount. The nicotine formulation is then delivered to the patientover a second period of time in a second dosage amount. The first andsecond periods of time may be any period of time including indefinitely,but is more typically between one week and two months, preferably is atime period within 1, 2, 3, 4 or 5 weeks. The second dosage amount mayalso vary, being greater or less than the first dosage amount, dependingupon the patient's reaction to the first dose. In all methods dosageamounts may be ramped up from a low amount to a higher amount to adjustfor patient sensitivity to the nicotine formulations of the invention.Thus a novel aspect of this method is to start the sensitive patientwith a lower dosage formulation that allows the patient to use less of atobacco product as a source of nicotine. As the patient becomesaccustomed to using the nicotine formulations of the invention, thedosage amount may be increased thus allowing a reduction in tobacco useas a source of nicotine. In this manner the patient can be freed fromthe health hazards of tobacco quickly and efficiently without sufferingthe effects of nicotine withdrawal. Once the formulas of the inventionhave entirely replaced tobacco as a source of nicotine, the dosageamount of the formulations of the invention may be gradually reduced toaddress the nicotine addition.

In another embodiment of the invention the different groups of targetscan be designed to target different groups of areas of the lung. Thus,for example, as shown in Table 1 the areas of the lung are broken downinto six general areas, these six general areas or even three generalareas could be targeted (See Table 1). The higher levels of therespiratory tract can be targeted using larger and larger particles.

TABLE 1 Subdivision of the Respiratory Tree Generation Name  0 Trachea 1 Primary bronchi  2 Lobar bronchi  3 Segmental bronchi  4 Subsegmentalbronchi  5 Small bronchi ↓ 10 11 Bronchioles, primary and secondary ↓ 1314 Terminal bronchioles ↓ 15 16 Respiratory bronchioles ↓ 18 19 Alveolarducts ↓ 23 24 Alveoli

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 compares the arterial nicotine profiles produced for cigarettesand various nicotine replacement therapies. The data is adapted fromRigotti, N. A., NEJM vol. 346, No. 7, (February 2002).

FIG. 2 depicts the mean arterial plasma nicotine concentrations for 16human patients.

FIG. 3 depicts the mean craving scores for 16 human patients.

FIG. 4 summarizes the modified Fagerström test for evaluating intensityof physical dependence on nicotine. Adapted with permission fromHeatherton T F, Kozlowski L T, Frecker R C, Fagerström K O. TheFagerström test for nicotine dependence: a revision of the FagerströmTolerance Questionnaire. Br J Addict 1991; 86:1119-27.

DEFINITIONS

Unless defined otherwise, all technical and scientific terms used hereinhave the meaning commonly understood by a person skilled in the art towhich this invention belongs. The following references provide one ofskill with a general definition of many of the terms used in thisinvention: Singleton et al., Dictionary of Microbiology and MolecularBiology (2nd ed. 1994); The Cambridge Dictionary of Science andTechnology (Walker ed., 1988); The Glossary of Genetics, 5th Ed., R.Rieger et al. (eds.), Springer Verlag (1991); and Hale & Marham, TheHarper Collins Dictionary of Biology (1991). As used herein, thefollowing terms have the meanings ascribed to them unless specifiedotherwise.

An “antidepressant” refers to a substance that is used in the treatmentof mood disorders, as characterized by various manic or depressiveaffects.

The term “anxiolytic” refers to any compound that has the effect ofrelieving anxiety.

An “aerosol” is a cloud of solid or liquid particles suspended in a gas.The particles may be formed from any suitable composition including, butnot limited to a solid such as a powder, a liquid, a gel, a cream, asuspension, an emulsion, or a colloidal mixture. Alternatively, anysemi-solid or semi-liquid may be used. Methods of forming aerosols fromprepared compositions are well-known in the art and described hereinbelow.

A “bioadhesive component” is one which aids the compound containing itin associating with biological tissue.

A “slow release component” is one which imparts the ability to dissolve,be absorbed, transported or broken down more slowly thereby allowing thecompound containing the slow release formula to persist.

When nicotine enters the circulatory system of a human patient it isoxidized to cotinine within four to six hours. The present inventionincludes the administration of cotinine and other nicotine derivativesprovided such derivatives do not result in unacceptable adverse effects

The term “nicotine” is intended to mean the naturally occurring alkaloidknown as nicotine, having the chemical nameS-3-(1-methyl-2-pyrrolidinyl)pyridine, which may be isolated andpurified from nature or synthetically produced in any manner. This termis also intended to encompass the commonly occurring salts containingpharmacologically acceptable anions, such as hydrochloride,hydrobromide, hydroiodide, nitrate, sulfate or bisulfate, phosphate oracid phosphate, acetate, lactate, citrate or acid citrate, tartrate orbitartrate, succinate, maleate, fumarate, gluconate, saccharate,benzoate, methanesulfonate, ethanesulfonate, benzenesulfonate, p-toluenesulfonate, camphorate and pamoate salts. Nicotine is a colorless to paleyellow, strongly alkaline, oily, volatile, hygroscopic liquid having amolecular weight of 162.23 and the formula:

Structure and ionization of nicotine. Nicotine is approximately 10% ofthe particulate weight in cigarette smoke. Brand differences change thispercentage. It is monoprotonated at most physiological pH values. Thediprotonated ion would exist at pH values found in the stomach.Metabolism is largely due to oxidation. Cotinine is a major metabolite;however, there are at least 4 primary metabolites of nicotine and allare encompassed by the use of this term herein.

The term “form of nicotine” further includes any pharmacologicallyacceptable derivative, metabolite or analog of nicotine which exhibitspharmacotherapeutic properties similar to nicotine. Such derivatives andmetabolites are known in the art, and include cotinine, norcotinine,nornicotine, nicotine N-oxide, cotinine N-oxide, 3-hydroxycotinine and5-hydroxycotinine or pharmaceutically acceptable salts thereof. A numberof useful derivatives of nicotine are disclosed within the Physician'sDesk Reference (most recent edition) as well as Harrison's Principles ofInternal Medicine. In addition, applicants refer to U.S. Pat. Nos.5,776,957; 4,965,074; 5,278,176; 5,276,043; 5,227,391; 5,214,060;5,242,934; 5,223,497; 5,278,045; 5,232,933; 5,138,062; 4,966,916;4,442,292; 4,321,387; 5,069,094; 5,721,257; all of which areincorporated herein by reference to disclose and describe nicotinederivatives and formulations.

“Free base nicotine” refers to the form of nicotine that predominates athigh pH levels. Free base nicotine is particularly potent and moreaddictive than nicotine salts which display a lower affinity tonicotinic receptors.

“A pharmaceutically active nicotine formulation” is a formulation havingat least two forms of nicotine as components, and may include additionaladditives and drug dosages.

The physiologically active form of nicotine is the S-(−)-isomer. Certaincompounds of the present invention may exist in particular geometric orstereoisomeric forms. The present invention contemplates all suchcompounds, including cis and trans isomers, R and S enantiomers,diastereomers, the racemic mixtures thereof, and other mixtures thereof,as falling within the scope of the invention. Additional asymmetriccarbon atoms may be present in a substituent such as an alkyl group. Allsuch isomers, as well as mixtures thereof, are intended to be includedin this invention.

The term “dual-release” is used herein to refer to a formulationcomprised of two components, one which releases nicotine or a nicotinederivative or nicotine substitute immediately, and one component whichreleases nicotine or a nicotine derivative or nicotine substitute over aprolonged period of time.

The term “diameter” is used herein to refer to particle size as given inthe “aerodynamic” size of the particle. The aerodynamic diameter is ameasurement of a particle of unit density that has the same terminalsedimentation velocity in air under normal atmospheric conditions as theparticle in question. In connection with the present invention, it isimportant that particles, on average, have the desired diameter so thatthe particles can be inhaled and targeted to a specific area of thelungs. To target the alveolar ducts and alveoli the particles shouldhave a diameter in a range of about 0.5μ to about 2μ.

The term “porous membrane” shall be interpreted to mean a membrane ofmaterial in the shape of a sheet having any given outer perimeter shape,but preferably covering a package opening which is in the form of anelongated rectangle, wherein the sheet has a plurality of openingstherein, which openings may be placed in a regular or irregular pattern,and which openings have a diameter in the range of 0.25μ to 4μ and apore density in the range of 1×10⁴ to about 1×10⁸ pores per squarecentimeter. The membrane functions to form an aerosolized mist when theformulation is forced through it. Those skilled in the art maycontemplate other materials which achieve this function as suchmaterials are intended to be encompassed by this invention.

The terms “treatment”, “treating”, and the like are used interchangeablyherein to generally mean obtaining a desired pharmacological and/orphysiological effect. The terms are used in a manner somewhatdifferently than the terms are typically used in that what is intendedby the method of treatment of the invention is to allow a patient toovercome an addiction to nicotine and thereby allow the patient to quitsmoking. The treating effect of the invention provides a psychologicaleffect in that the invention originally delivers high doses of nicotinein a manner that simulates the nicotine delivery obtained from acigarette. The patient then becomes accustomed to relying on themethodology of the invention to provide an immediate “rush” of nicotine.Thereafter, the particles of the aerosol are made larger. This preventsthe particles from penetrating deeply into the lung and, therefore, tosome extent, diminishes the “rush” of nicotine. However, the same amountof nicotine is still given to the patient in order to satisfy theoverall nicotine craving. Eventually, the treatment of the inventionreduces the amount of nicotine so as to allow the patient to completely“wean” off of nicotine and to quit smoking.

All publications mentioned herein are incorporated herein by referenceto described and disclose specific information for which the referencewas cited in connection with. The publications discussed herein areprovided solely for their stated disclosure prior to the filing date ofthe present application. Nothing herein is to be construed as anadmission that the invention is not entitled to antedate suchpublications by virtue of prior invention. Further, the actualpublication date may be different from that stated on the publicationand as such may require independent verification of the actualpublication dates.

DETAILED DESCRIPTION OF EXEMPLARY EMBODIMENTS OF THE INVENTION I.Introduction

The present invention provides a novel formulations and methods fortheir use that pharmacologically mimic the delivery of nicotine producedby smoking a cigarette. These formulations may be delivered in a singledose, such as a single breath from an inhaler, and provide an effectivemeans for addressing and potentially eliminating a person's addiction totobacco products including those used for smoking, chewing and sniffing.This nicotine dosage is completely tobacco-free and thus provides thepatient an opportunity to address tobacco use and an addiction tonicotine individually. The separation of the habitual use of tobacco andnicotine addiction eases withdrawal from the dangerous habit and isbelieved to increase the opportunity to succeed in breaking the habit.

The formulations of the claimed invention contain at least two forms ofnicotine that in combination mimic the pharmacological delivery ofnicotine produced by smoking a cigarette without exposing the user totobacco products. The formulas of the invention may be administered inany suitable manner known in the art that allows the pharmacologicaldosage pattern for nicotine described herein for the invention.Typically delivery will be by inhalation or sniffing the product intothe respiratory tract, including the deep lung alveoli. In this mannerthe invention provides a nicotine dosage that rapidly peaks and thentrails off maintaining arterial nicotine concentrations that mirrorthose produced by smoking tobacco (FIG. 1). The invention produces thenicotine dosage pattern by providing at least two forms of nicotine. Thefirst form of nicotine includes sufficiently small particles that may beinhaled deeply into the lung, i.e. 50% or more of the particles areinhaled deeply into the lung and thereby quickly enter the patient'scirculatory system. The invention also provides a second form ofnicotine that is in a slow release form. This second form of nicotineensures that the patient's arterial plasma nicotine concentration ismaintained at levels that minimize craving for nicotine over a moreextended time period.

The two forms of nicotine are typically dispersed in the form ofparticles that may originate from dry powder, liquid suspension oremulsion, microspheres suspended in an aqueous solution or dried, or anyother physical manifestation that can be aerosolized allowing theparticles to reach the intended region of the lung. By controlling thephysical and chemical characteristics of the particles the release ofthe nicotine formulation to a patient's circulatory system as describedherein may be controlled. The first and second forms of nicotine may bepresent in the formulations of the invention in different physicalforms. For example, the formulation of the invention may be a liquidsuspension of microspheres where the first form of nicotine is dissolvedin the fluid component and the second form of nicotine is encapsulatedin the microshperes. Alternatively, the formulation could be a drypowder where the first and second forms of nicotine are distinguished byparticle size, roughness, diameter, composition or any combination ofdifferences. A third exemplary formulation would be a heterogeneoussuspension of microspheres where the first and second forms of nicotineare encapsulated in separate microsphere populations.

Particles suitable for use in the instant invention may be fabricatedwith the appropriate material, surface roughness, diameter and densityfor localized delivery to selected regions of the respiratory tract suchas the deep lung, central or upper airways. For example, to increase theaerodynamic diameter, higher density or larger particles may be used forupper airway delivery or, smaller or lower density, e.g., porousparticles, may be utilized for deep lung deposition. More preferably amixture of different sized particles in a formulation may beadministered to target different regions of the lung in oneadministration. Particles with degradation and release times rangingfrom seconds to hours can be designed and fabricated, based on factorssuch as the particle material. Techniques for fabricating such particlesare well known in the art. For example, particles of the invention maybe a form of nicotine that is a dry powder manufactured from nicotinewith additional, optional, materials added to the formulation to impartdesired characteristics as described in more detail below.

The present invention is also advantageous in that the rate at which thedelivered nicotine enters the circulatory system can be graduallymodulated, for example by gradually increasing the size of theaerosolized particles delivered to the patient leading to deposition inthe parts of the respiratory tract from which absorption is slower. Thiscan be done over any desired period of time and in any desired number ofphases.

Moreover, the invention provides a means whereby the amount of nicotinedelivered to the patient may be gradually decreased in a number ofdifferent ways. For example, nicotine delivery may be decreased bydecreasing the concentration of nicotine in the tobacco-freeformulation; nicotine may be decreased by decreasing the number of dosestaken by the patient over a given period of time; nicotine may also bedecreased by decreasing the size of the dose administered to thepatient; and finally, nicotine delivery may be decreased by altering thenature of the formulation, as described herein.

As depicted in FIG. 1, current nicotine therapies are characterized byslow absorption and low blood levels of nicotine, limiting theirutility. The present invention replaces the nicotine that a patientreceives from using a tobacco product by providing a rapid pulse ofbioavailable nicotine to the patient, followed by a slow release ofnicotine providing a prolonged circulating concentration of nicotine.More specifically, the present invention provides a treatmentmethodology wherein a patient's initial arterial nicotine plasmaconcentration over a selected time, i.e., the arterial nicotine plasmaconcentration-rate profile, substantially correlates to that of thepatient when smoking a cigarette; the slow release component of theformulation then maintains a minimum level of circulating nicotine overa longer period of time, in the range of 1 to 24 hours.

One treatment methodology of the present invention creates an aerosol ofnicotine particles. As noted previously, the nicotine particles may bein powder form, or formed initially as droplets from any liquidcontaining nicotine including a solution or suspension of nicotine andaerosolized in any known manner including (1) moving the formulationthrough a porous membrane in order to create particles or (2) a drypowder where the particles of powder have been designed to have adesired diameter. By increasing the size of the particles from about 1-2microns (μm) upwards causes the particles to be deposited higher in therespiratory tract. Without limiting the scope of the invention, it isgenerally known that higher regions of the respiratory tract have lesstissue surface area than lower regions. As the rate of particleabsorption is known to be directly proportional to the surface area ofthe tissue on which the particles are deposited, nicotine is absorbedmore slowly through the mucosal membranes of the upper respiratorytract. Thus the effect of increasing particle size is to deposit theinhaled particle in a higher region of the respiratory tract withconcomitant reduced absorption rate over time and a more sustained drugprofile. Of course other mechanisms may also play a significant role inthe release of nicotine to the circulatory system, and the presentinvention does not exclude such mechanisms. For example, clearance oflarger particles from the upper respiratory tract may result intransport of those forms of nicotine to alternative locations and/or maycontribute to the delay or sustained release of the nicotine form to thecirculatory system.

Thus one method of practicing the present invention is to provide aformulation comprising two forms of nicotine, a first form characterizedby fine particles of small diameter and a second form characterized bylarger particles. The larger particles deposit in the upper respiratorytract providing low level sustained drug release, while the smallerparticles penetrate to the deep lung providing a rapid pulse ofavailable nicotine similar to that provided by a cigarette.

The method of the invention has applicability to smokers and users ofother tobacco products wishing to quit or trying to quit who haveexperienced all or any of the nicotine withdrawal symptoms associatedwith withdrawal from tobacco products. These symptoms include cravingfor nicotine, irritability, frustration or anger, anxiety, drowsiness,sleep disturbances, impaired concentration, nervousness, restlessness,decreased heart rate, increased appetite, and weight gain among others.

While particularly applicable to addressing habitual use of tobaccoproducts, pulmonary, oral, or parenteral administration of nicotinecould be of value for the treatment of other diseases, such as forpatients suffering from neurodegenerative diseases, psychiatricdisorders and other central nervous system disorders responsive tonicotinic receptor modulation (see U.S. Pat. Nos. 5,187,169; 5,227,391;5,272,155; 5,276,043; 5,278,176; 5,691,365; 5,885,998; 5,889,029;5,914,328). Such diseases include, but are not limited to, seniledementia of the Alzheimer's type, Parkinson's disease, schizophrenia,obsessive-compulsive behavior, Tourette's Syndrome, depression,attention deficit disorder, myasthenia gravis and drug addiction. Theseembodiments and others are discussed in greater detail, below. SeeMasterson (1991) U.S. Pat. No. 5,069,904; Wesnes and Warburton (1984)Psychopharmacology 82:147-150; and Warburton et al. (1986)Psychopharmacology 89:55-59.

II. Tobacco-Less formulations

Tobacco-less formulations of the present invention are preferablysuitable for formation of aerosols containing at least two forms ofnicotine. Preferable embodiments are powders, liquids, emulsions, andsuspensions (e.g., suspensions of microspheres). The formulations mayoptionally include other drugs, excipients, permeation enhancers,preservatives, absorption enhancers, binding agents, buffers, and thelike that enhance the efficacy or ease the use of the claimed invention.Typical nicotine forms of the invention include nicotine dissolved inwater or dry powder nicotine with a carrier used to adjust the pH to thedesired range. Methods of formulating liquids and liquid inhalers aredisclosed in U.S. Pat. Nos. 5,364,838; 5,709,202; 5,497,763; 5,544,646;5,718,222; 5,660,166; 5,823,178; and 5,910,301; all of which areincorporated by reference to describe and disclose such. Contemplatedcomponents of the claimed invention are discussed in greater detail,below.

Powder or granular forms of the invention may be combined with asolution and with a diluting, dispersing or surface-active agent.Additional preferred compositions for administration include abioadhesive to retain the agent at the site of administration; a spray,paint, or swab applied to the mucosa or epithelium; a slow dissolvingpill or lozenge, or the like. The composition may also be in the form oflyophilized powder, which can be converted into solution, suspension, oremulsion before administration. The formulations of the invention arepreferably sterilized and stored in unit-dose or multi-dose containerssuch as sealed vials or ampoules using methods well-known to those ofskill in the art.

A. Suitable Forms of Nicotine

Formulations of the present invention include two forms of nicotine thatin combination mimic the pharmacological profile of nicotine deliveryproduced by a cigarette. The nicotine forms of the invention may bepowders, liquids, or encapsulated. Preferably the nicotine forms aresuitable for formation of aerosols that are amenable to inhalation. Thepreparation is such that the inhaled nicotine will be both in a formthat provides rapid absorption and also in a form that provides a moresustained rate of absorption. For example, when the claimed formulationis inhaled, the first form of nicotine has a smaller particle diameterthan the second form of nicotine. This allows the first form of nicotineto be deposited in the deep lung where it is rapidly transferred to theuser's blood stream and reaches the users central nervous system within5 minutes, preferably in less than 4, 3, 2 or 1 minute. The largerparticle size of the second form of nicotine results in deposition ofthis nicotine form higher up in the respiratory tract. As a result, thesecond form of nicotine is released more slowly to the users circulatorysystem with a more sustained effect. Alternatively the treatment is amixture of immediate and slow release forms of nicotine. Nicotine formsof the invention are discussed in greater detail, below.

1. First Form of Nicotine

The first form of nicotine is preferentially inhaled as this method ofadministration provides the most rapid delivery without resorting toinvasive techniques such as injection. Inhalation allows for a suitablefirst form of nicotine arterial concentration in the patient within 5minutes of delivery. Typically this arterial concentration is at least10, 12, 14 or 15 ng/ml, and this concentration is achieved within 5,preferably within 4, 3, 2, or 1 minute or less from inhalation of theclaimed formulation.

To facilitate the rapid delivery of the drug to the user's centralnervous system when inhaled, the particle or droplet size of the firstform of nicotine is controlled and kept small in order to allow theparticles to reach the deep lung. Typically this size is between about 1μm and about 4 μm in diameter, more preferably about 2 or 3 μm.

The first form of nicotine may have a fluid component having a basic pH,preferably having a pH of more than 7.5, 8.0, or 8.5. A basic pHfacilitates formation of the more potent free base form of nicotine. Asdiscussed below, the nicotine forms of the claimed formulation may beencapsulated for example in microspheres. Encapsulation allows thenicotine forms of the formulation to be segregated and therefore theymay be delivered with different additives, including buffers adjustingpH, due to their respective microenvironments.

2. Second Form of Nicotine

The second form of nicotine in the formulations of the invention arepresent in an amount to maintain a second form of nicotine arterialconcentration in the patient for at least 60 minutes after delivery.This second form of nicotine formulation, if administered on its own,would lead to an arterial concentration that is generally lower than thefirst form of nicotine arterial concentration, typically being at leastabout 8 ng/ml, preferably about 6 ng/ml, more preferably at least about5 ng/ml, or at least about 4, 3, 2 ng/ml.

Delivery of the second form of nicotine may be performed using anysuitable method with preferable methods being buccally (e.g., as a gum,quick dissolve strip, or lozenge composition), transdermal patch,inhalation, or other method that allows for sustained release of thesecond form of nicotine over a period of several minutes to hours,preferably at least 30, 40, or 60 minutes, more preferably 90 or 120minutes. The second form of nicotine may be delivered at any pH, but ismore preferably delivered at a pH which is most suitable for aparticular delivery route.

A preferred method of administering the formulations of the invention isthrough inhalation. When inhaled, the second form of nicotine may have alarger particle size than the first form of nicotine. As discussedelsewhere in this specification, the larger particle size results in thesecond form of nicotine being deposited preferentially in the upperrespiratory tract rather than the deep lung. Deposition in the higherrespiratory airways results in the second form of nicotine reaching theblood system and the receptors of the patient's central nervous systemmore slowly than is the case for the first form of nicotine deposited inthe deep lung. This aids in the sustained release of lower levels of thesecond form of nicotine to the blood as desired in mimicking thepharmacological administration of nicotine via a cigarette. Thusparticles or droplets of the formulation containing the second form ofnicotine are preferably in the range between about 4 μm and about 12 μm,more preferably between about 5 μm and about 10 μm, preferentiallybetween about 6 μm and about 8 μm in diameter, as these sizes facilitatedeposition of the particles or droplets in the upper airway passages ofthe lung. It is also possible to slow the absorption using a sustainedrelease formulation.

It is also possible to deliver the second form into the deep lung andother parts of the respiratory tract and provide prolonged elevatedlevels of nicotine in the arterial blood supply through the sustainedrelease of the second form of nicotine from slow release formulationsthat are reside over a suitable period of time in the respiratory tract.The residence time may be prolonged by deep lung delivery, or throughthe use of bioadhesive components. The component of the formulation mayoptionally include a slow release component such as liposomes or otherencapsulating materials well known to those of skill in the artincluding packaging within microspheres. Encapsulation in microsphereshas the added advantage of facilitating delivery of the first and secondforms of nicotine at different pH values. For example, the first form ofnicotine may be delivered in free base form having a basic pH whereasthe second form of nicotine is delivered in salt form as an acidic pH.As is known, the free base form interacts with the nicotinic receptoreliciting a larger response than more acidic? forms of the drug.

Preferred microspheres for use in the invention include polyglycolidemicrospheres. Microspheres may also optionally include a bioadhesivecomponent such as hyluronic acid.

Microsomes and liposomes of the present invention may be constructedusing techniques well-known to those of skill in the art. For example,liposomes containing the second form of nicotine of the presentinvention may be prepared by suspending a thin layer of purifiedphospholipids in a solution containing the second form of nicotine andthen treating the suspension in a conventional manner such asultrasonication. A “Liposome” is a closed vesicle of lipid bilayerencapsulating an aqueous compartment therein. It is known that the lipidbilayer membrane structure is extremely similar to biological membranes.

3. Preparing Nicotine Particles

Analysis of Nicotine Containing Aerosols

Purity of a nicotine-containing aerosol may be determined using a numberof methods, examples such as described in Sekine et al., Journal ofForensic Science 32:1271 1280 (1987) and Martin et al., Journal ofAnalytic Toxicology 13:158 162 (1989). One method involves forming theaerosol in a device through which a gas flow (e.g., air flow) ismaintained, generally at a rate between 0.4 and 60 L/min. The gas flowcarries the aerosol into one or more traps. After isolation from thetrap, the aerosol is subjected to an analytical technique, such as gasor liquid chromatography that permits a determination of compositionpurity.

A variety of different traps are used for aerosol collection. Thefollowing list contains examples of such traps: filters; glass wool;impingers; solvent traps, such as dry ice-cooled ethanol, methanol,acetone and dichloromethane traps at various pH values; syringes thatsample the aerosol; empty, low-pressure (e.g., vacuum) containers intowhich the aerosol is drawn; and, empty containers that fully surroundand enclose the aerosol generating device. Where a solid such as glasswool is used, it is typically extracted with a solvent such as ethanol.The solvent extract is subjected to analysis rather than the solid(i.e., glass wool) itself. Where a syringe or container is used, thecontainer is similarly extracted with a solvent.

The gas or liquid chromatograph discussed above contains a detectionsystem (i.e., detector). Such detection systems are well known in theart and include, for example, flame ionization, photon absorption andmass spectrometry detectors. An advantage of a mass spectrometrydetector is that it can be used to determine the structure of opioiddegradation products.

Particle size and composition of the different forms of nicotine of theformulation may be controlled using techniques well-known to those ofskill in the art.

Particle size distribution of aerosols produced using formulations ofthe invention may be determined using any suitable method in the art(e.g., cascade impaction). An Andersen Eight Stage Non-viable CascadeImpactor (Andersen Instruments, Smyrna, Ga.) linked to a source ofaerosol by a mock throat (USP throat, Andersen Instruments, Smyrna, Ga.)is one system used for cascade impaction studies.

Inhalable aerosol mass density may be determined, for example, bydelivering a drug-containing aerosol into a confined chamber via aninhalation device and measuring the mass collected in the chamber.Typically, the aerosol is drawn into the chamber by having a pressuregradient between the device and the chamber, wherein the chamber is atlower pressure than the device. The volume of the chamber shouldapproximate the tidal volume of an inhaling patient.

Inhalable aerosol nicotine mass density is determined, for example, bydelivering an aerosol of the invention into a confined chamber via aninhalation device and measuring the amount of active drug compoundcollected in the chamber. Typically, the aerosol is drawn into thechamber by having a pressure gradient between the device and thechamber, wherein the chamber is at lower pressure than the device. Thevolume of the chamber should approximate the tidal volume of an inhalingpatient. The amount of nicotine collected in the chamber is determinedby extracting the chamber, conducting chromatographic analysis of theextract and comparing the results of the chromatographic analysis tothose of a standard containing known amounts of drug.

Inhalable aerosol particle density is determined, for example, bydelivering an aerosol of the invention into a confined chamber via aninhalation device and measuring the number of particles of given sizecollected in the chamber. The number of particles of a given size may bedirectly measured based on the light-scattering properties of theparticles. Alternatively, the number of particles of a given size isdetermined by measuring the mass of particles within the given sizerange and calculating the number of particles based on the mass asfollows: Total number of particles=Sum (from size range 1 to size rangeN) of number of particles in each size range. Number of particles in agiven size range=Mass in the size range/Mass of a typical particle inthe size range. Mass of a typical particle in a given sizerange=π*D³*θ/6, where D is a typical particle diameter in the size range(generally, the mean boundary MMADs defining the size range) in microns,θ is the particle density (in g/mL) and mass is given in units ofpicograms (g⁻¹²).

Rate of inhalable aerosol particle formation is determined, for example,by delivering aerosol into a confined chamber via an inhalation device.The delivery is for a set period of time (e.g., 3 s), and the number ofparticles of a given size collected in the chamber is determined asoutlined above. The rate of particle formation is equal to the number of100 nm to 5 μm particles collected divided by the duration of thecollection time.

Rate of aerosol formation is determined, for example, by deliveringaerosol phase drug into a confined chamber via an inhalation device. Thedelivery is for a set period of time (e.g., 3 s), and the mass ofparticulate matter collected is determined by weighing the confinedchamber before and after the delivery of the particulate matter. Therate of aerosol formation is equal to the increase in mass in thechamber divided by the duration of the collection time. Alternatively,where a change in mass of the delivery device or component thereof canonly occur through release of the aerosol phase particulate matter, themass of particulate matter may be equated with the mass lost from thedevice or component during the delivery of the aerosol. In this case,the rate of aerosol formation is equal to the decrease in mass of thedevice or component during the delivery event divided by the duration ofthe delivery event.

Dry Powder Formulations

Methods for producing dry powder formulations with particle sizeslimited to an inhalable aerodynamic are well known by those of skill inthe art. They include but are not limited to, milling, spray-drying,freeze-drying, lyophilization, absorption and adsorption of activeingredients into and onto carrier particles.

Dry powder formulations obtainable according to the invention mayinclude a pharmaceutically inactive carrier of noninhalable particlesize, a finely divided pharmaceutically active compound of inhalableparticle size and to improve the resistance to moisture—magnesiumstearate, and they are preferably present in the form of “interactive(or ordered or adhesive) mixtures”. If desired, the dry powderformulations can also contain a proportion of carrier material ofinhalable particle size. In principle, the constituents can be mixedwith one another in any desired sequence, where, however, mixing shouldexpediently be carried out in such a way that the particles of theconstituents are essentially retained as such, i.e. are not destroyed,for example, by granulation and the like. Mixing can be carried out in amanner known per se, for example in a tumble mixer.

The expression “interactive mixture” or “ordered mixture” or “adhesivemixture” is familiar to the person skilled in the art and in the contextof the present invention comprises dry powder formulations in which thepharmacologically inactive carrier is present in a particle size whichis noninhalable or mainly noninhalable, and in which microfine particlesof the nicotine forms are bound to the carrier particles by adhesion(i.e. are not contained in the carrier, e.g. in the form of granules).

The amount of nicotine in the formulations obtainable according to theinvention may vary within wide ranges and is to a high extent dependenton the particular nicotine form and up to a certain degree also on thepowder inhaler used. Typically, the nicotine concentration can beapproximately 0.1 to 10% by weight, in particular approximately 0.1 to5% by weight, based on the total formulation. Occasionally, higher orlower concentrations can also be expedient however active compoundconcentrations of below 0.001% by weight or below 0.01% by weight rarelyoccur.

Microsphere Formulations

Preparation of microspheres, including liposomes is well known in theart, as are compositions providing microspheres with differentdissolution rates. Thus formulations of the invention may includelipophilic substances that can enhance absorption of the agent throughthe mucosa or epithelium of the nasal cavity. The forms of nicotine ofthe invention may be mixed with a lipophilic adjuvant alone or incombination with a carrier, or may be combined with one or several typesof micelle or liposome substances. Among the preferred lipophilicsubstances are cationic liposomes included of one or more of thefollowing: phosphatidyl choline, lipofectin, DOTAP, a lipid-peptoidconjugate, a synthetic phospholipid such as phosphatidyl lysine, or thelike. These liposomes may include other lipophilic substances such asgangliosides and phosphatidylserine (PS). Also preferred are micellaradditives such as GM-1 gangliosides and phosphatidylserine (PS), whichmay be combined with the agent either alone or in combination. GM-1ganglioside can be included at 1-10 mole percent in any liposomalcompositions or in higher amounts in micellar structures. Protein agentscan be either encapsulated in particulate structures or incorporated aspart of the hydrophobic portion of the structure depending on thehydrophobicity of the active agent.

For one skilled in the art, the release rate from microspheres can beeasily modified ranging from days to months by altering the ratio of thecopolymers. For example, the Eligard product uses the ATRIGEL® DeliverySystem, a polymeric (non-gelatin containing) delivery system consistingof a biodegradable poly(DL-lactide-co-glycolide) (PLGH) polymerformulation dissolved in a biocompatible solvent, N-methyl-2-pyrrolidone(NMP). The leuprolide delivery rate is described by a one-month releaseby using co-polymer with a 50:50 molar ratio of DL-lactide to glycolidecontaining carboxyl end groups. In the 3 and 6-month product, theleuprolide delivery rate is achieved by using co-polymer with a 75:25molar ratio of DL-lactide to glycolide with hexanediol or an 85:15 molarratio of DL-lactide to glycolide with hexanediol, respectively. Clearly,the greater the ratio of PLA to PGA the more prolonged the release.

Examples of commercially available peptide/protein controlled, releasesystems based on PLGA include:

Drug Trade name Company Polymer Route Application buserelinProfact□Depot, Hoechst PLGA s/c implant Prostate acetate Suprefact□DepotMarion cancer Roussel goserelin Zoladex□Depot Astra Zeneca PLGA s/cimplant Prostate acetate cancer, endometrioses Leuprolide EligardSanofi- 1, 3, 4 and 6 Prostate acetate aventis month cancer suspensioninjection leuprorelin Lupron□Depot, Takeda- PLGA 3-month Prostateacetate Enantone□Depot, Abbott PLA depot cancer, Enantone□Gynsuspension, endometrioses Depot 1-month Trenantone□ suspension 3-monthsuspension octreotide Sandostatin Novartis PLGA s/c GH acetate LAR□DepotPharma suspension suppression, anti cancer triptorelin Decapeptyl ®Debiopharma PLGA s/c depot LHRH agonist, Depot injection prostate cancerrecombinant Nutropin□Depot, Genentech- PLGA monthly s/c Growth human[discontinued Alkermes injection hormone growth commercialisationdeficiency hormone since June 2004]

One preferred liposomal formulation employs Depofoam. An agent can beencapsulated in multivesicular liposomes, as disclosed in the copendingapplication entitled “High and Low Load Formulations of IGF-I inMultivesicular Liposomes,” U.S. patent application Ser. No. 08/925,531,filed Sep. 8, 1997, herein incorporated by reference. The mean residencetime of agent at the site of administration can be prolonged with aDepofoam composition.

4. Supplemental Drugs

Methods for formulating pharmaceutical compositions are generally knownin the art. A thorough discussion of formulation and selection ofpharmaceutically acceptable carriers, stabilizers, and isomolytes can befound in Remington's Pharmaceutical Sciences (18.sup.th ed.; MackPublishing Company, Eaton, Pa., 1990), herein incorporated by reference.

In addition to the nicotine forms discussed above, the tobacco-lesscompositions of the present invention may optionally includesupplemental pharmaceutically-active components. These supplementalcomponents may aid in delivery of the nicotine forms of the formulation,provide further support of the patient's program to terminate theirnicotine addiction, treat diseases, or make the formulations of theinvention more acceptable to the patient-user.

Particularly preferred supplemental drugs include antidepressants andanxiolytics such as selective serotonin reuptake inhibitors, e.g.,citalopram, escitalopram, fluoxetine, paroxetine, sertraline, and thelike. Serotonin and norepinephrine reuptake inhibitors are alsopreferred, such as duloxetine, venlafaxine, and the like. Norepinephrineand dopamine reuptake inhibitors such as bupropion may also be used.Tetracyclic antidepressants such as mirtazapine; combined reuptakeinhibitors and receptor blockers such as trazodone, nefazodone,maprotiline; tricyclic antidepressants, such as amitriptyline,amoxapine, desipramine, doxepin, imipramine, nortriptyline,protriptyline and trimipramine; monoamine oxidase inhibitors, such asphenelzine, tranylcypromine, isocarboxazid, selegiline; benzodiazepinessuch as lorazepam, clonazepam, alprazolam, and diazepam; serotonin 1Areceptor agonists such as buspirone, aripiprazole, quetiapine,tandospirone and bifeprunox; and a beta-adrenergic receptor blocker,such as propranolol may also be added to enhance the claimedtobacco-less formulations of the present invention.

The formulations of the present invention may also optionally includeother pharmacologic agents such as UTP, amiloride, antibiotics,bronchodilators, anti-inflammatory agents, and mucolytics (e.g.n-acetyl-cysteine). In addition to including other therapeutic agents inthe formulation itself, the formulations of the present invention mayalso be administered sequentially or concurrently with the one or moreother pharmacologic agents identified herein. The amounts of formulationand pharmacologic agent depend, for example, on what type ofpharmacologic agent(s) are used, and the scheduling and routes ofadministration

Supplemental drugs may be delivered concomitantly with the formulationsof the present invention, or may be administered independently.Supplemental drug delivery may be via any suitable method known in theart including orally, inhalation, injection, etc.

B. Pharmaceutically Acceptable Excipients

The formulations of the present invention are administered to a humanand may contain one or more pharmaceutically-acceptable excipients, orcarriers. Suitable excipients and their formulations are described inRemington's Pharmaceutical Sciences, 16th ed., 1980, Mack PublishingCo., edited by Oslo et al.

For the exact volumetric dosage of the formulations of the invention,dilution of the active compound with a pharmaceutically inactiveexcipient may be necessary in order to obtain a dosable unit amountmeeting the demands on dosage accuracy. Where necessary the dilution ischosen such that the amount applied from the inhaler exactly containsthe desired dose. The pharmacologically inactive excipient preferablyserves not only for dilution, but also for the adjustment of aflowability of the powder mixture or aerosol mist. The proportion ofcarrier material in the formulations obtainable according to theinvention can vary within a wide range depending on the dilution. Theproportion of carrier material to the total formulation can be, forexample, approximately 80 to 99.9% by weight, where, however, higher orlower proportions can also be advantageous depending on the nicotineform(s) of the formulation. The lower proportion of the excipient isadvantageous in order to minimize the possibility of adverse reactionsdue to the excipient, unless the excipient is known to be safe whendelivered by inhalation.

The carrier is preferably present in the formulation obtainableaccording to the invention in a particle size which is not inhalable.The carrier particles, however, should on the other hand not be toolarge, as this can have a disadvantageous effect on the FPF. The optimumparticle size of the carrier employed in this case as a rule depends onthe demands and specifications of the inhaler which is intended for theadministration of the formulation. In the context of the presentinvention, carriers having customary particle sizes can be used, andoptimum particle sizes can easily be determined from case to case by theperson skilled in the art. In general, however, the mean particlediameter (MMAD) of the carrier particles can be approximately 10 to 500μm and preferably approximately 50 to 200 μm.

Where applicable, the adhesion of the formulation particles to carrierparticles should be sufficient that no demixing takes place duringprocessing, transport, storage and dosage operations, but on the otherhand not so high that a detachment of the formulation particles which isas quantitative as possible is no longer guaranteed during thedispersion in the inhaler induced by the respiratory flow of thepatient. The effectiveness of the release of the active compoundparticles is especially dependent, in addition to the physicochemicalproperties of the active compound and the aerodynamic properties of thepowder inhaler, on the properties of the carrier, in particular thenature of the carrier and its surface structure, mean particle size andparticle size distribution.

In the context of the powder formulations of the present invention,fundamentally all carrier materials customarily used in dry powderformulations are suitable, for example mono- or disaccharides, such asglucose, lactose, lactose monohydrate, sucrose or trehalose, sugaralcohols, such as mannitol or xylitol, polylactic acid or cyclodextrin,glucose, trehalose and in particular lactose monohydrate in generalbeing preferred. If desired, the formulations can also contain two ormore carrier materials. If desired, in addition to noninhalable carrierparticles, the formulation can also contain a proportion of inhalablecarrier particles; for example in addition to relatively coarse lactosemonohydrate carrier particles it can contain a proportion of, forexample, 0.1 to 10% by weight of micronized lactose monohydrate, whichcan have, for example, a particle size diameter of at most 10 μm,preferably at most 5 μm, for at least 50% of the particles.

Water, saline, aqueous dextrose, and glycols are preferred liquidcarriers, particularly (when isotonic) for solutions. The carrier can beselected from various oils, including those of petroleum, animal,vegetable or synthetic origin, for example, peanut oil, soybean oil,mineral oil, sesame oil, and the like. Suitable pharmaceuticalexcipients include starch, cellulose, talc, glucose, lactose, sucrose,gelatin, malt, rice, flour, chalk, silica gel, magnesium stearate,sodium stearate, glycerol monostearate, sodium chloride, dried skimmilk, glycerol, propylene glycol, water, ethanol, and the like. Thecompositions can be subjected to conventional pharmaceutical expedients,such as sterilization, and can contain conventional pharmaceuticaladditives, such as preservatives, stabilizing agents, wetting, oremulsifying agents, salts for adjusting osmotic pressure, buffers, andthe like.

Other optional components of the formulations may include, but are notlimited to, buffers that enhance isotonicity such as water, saline,phosphate, citrate, succinate, acetic acid, and other organic acids ortheir salts. Typically, the pharmaceutically acceptable carrier alsoincludes one or more stabilizers, reducing agents, anti-oxidants and/oranti-oxidant chelating agents. The use of buffers, stabilizers, reducingagents, anti-oxidants and chelating agents in the preparation of proteinbased compositions, particularly pharmaceutical compositions, iswell-known in the art. See, Wang et al., “Review of Excipients and pHsfor Parenteral Products Used in the United States.” J. Parent. DrugAssn., 34(6):452-462 (1980); Wang et al., “Parenteral Formulations ofProteins and Peptides: Stability and Stabilizers,” J. Parent. Sci. andTech., 42:S4-S26 (Supplement 1988); Lachman, et al., “Antioxidants andChelating Agents as Stabilizers in Liquid Dosage Forms-Part 1,” Drug andCosmetic Industry, 102(1): 36-38, 40 and 146-148 (1968); Akers, M. J.,“Antioxidants in Pharmaceutical Products,” J. Parent. Sci. and Tech.,36(5):222-228 (1988); and Methods in Enzymology, Vol. XXV, Colowick andKaplan eds., “Reduction of Disulfide Bonds in Proteins withDithiothreitol,” by Konigsberg, pages 185-188.

Suitable buffers include acetate, adipate, benzoate, citrate, lactate,maleate, phosphate, tartarate, borate, tri(hydroxymethyl aminomethane),succinate, glycine, histidine, the salts of various amino acids, or thelike, or combinations thereof. See Wang (1980) at page 455. Suitablesalts and isotonicifiers include sodium chloride, dextrose, mannitol,sucrose, trehalose, or the like. Where the carrier is a liquid, it ispreferred that the carrier is hypotonic or isotonic with oral,conjunctival or dermal fluids and have a pH within the range of 4.5-8.5.Where the carrier is in powdered form, it is preferred that the carrieris also within an acceptable non-toxic pH range.

The formulations may also include an adjuvant such as cetyl trimethylammonium bromide, BDSA, cholate, deoxycholate, polysorbate 20 and 80,fusidic acid, or the like, and in the case of DNA delivery, preferably,a cationic lipid. Suitable sugars include glycerol, threose, glucose,galactose and mannitol, sorbitol. A suitable protein is human serumalbumin.

Preferred fluid compositions include one or more of a solubilityenhancing additive, preferably a cyclodextrin; a hydrophilic additive,preferably a mono or oligosachharide; an absorption promoting additives,preferably a cholate, a deoxycholate, a fusidic acid, or a chitosan; acationic surfactant, preferably a cetyl trimethyl ammonium bromide; aviscosity enhancing additive, preferably to promote residence time ofthe composition at the site of administration, preferably acarboxymethyl cellulose, a maltodextrin, an alginic acid, a hyaluronicacid, or a chondroitin sulfate; or a sustained release matrix,preferably a polyanhydride, a polyorthoester, a hydrogel, a particulateslow release depo system, preferably a polylactide co-glycolides (PLG),a depo foam, a starch microsphere, or a cellulose derived buccal system;a lipid based carrier, preferably an emulsion, a liposome, a niosomes,or a micelles. The composition can include a bilayer destabilizingadditive, preferably a phosphatidyl ethanolamine; a fusogenic additive,preferably a cholesterol hemisuccinate.

Pharmaceutically acceptable excipients may be volatile or nonvolatile.Volatile excipients, when heated, are concurrently volatilized,aerosolized and inhaled with the antihistamine. Classes of suchexcipients are known in the art and include, without limitation,gaseous, supercritical fluid, liquid and solid solvents. The followingis a list of exemplary carriers within the classes: water; terpenes,such as menthol; alcohols, such as ethanol, propylene glycol, glyceroland other similar alcohols; dimethylformamide; dimethylacetamide; wax;supercritical carbon dioxide; dry ice; and mixtures thereof.

These lists of carriers and additives is by no means complete and aworker skilled in the art can choose excipients from the GRAS (generallyregarded as safe) list of chemicals allowed in the pharmaceuticalpreparations and those that are currently allowed in topical andparenteral formulations.

C. Propellants

Tobacco-less formulations of the present invention may also include apropellant suitable for aerosolizing the pharmaceutically activenicotine formulation. Suitable propellants are well-known in the art andinclude compressed air, nitrogen, hydrofluoroalkanes (HFAs) and thelike. An important aspect of any propellant used in the presentinvention is that it not react with nicotine or otherpharmaceutically-active components of the tobacco-less formulations ofthe claimed invention.

III. Methodology

The penetration of aerosolized nicotine particles into the respiratorytract is determined largely by the size distribution of the particlesformed and may be also affected by the breathing pattern just prior to,during and just after the inhalation of the medication. The sites ofdeposition also depend on age and the pathophysiological condition ofthe person inhaling the medication. Under normal breathing conditions,larger particles, i.e., particles with a diameter greater than or equalto 5 μm, deposit predominantly on the upper airways of the lungs (seeFIG. 1). Particles having a diameter in a range of about >2 microns (μm)to <5 microns (μm) deposit predominantly in the central airways. Smallerparticles having a diameter □2 microns (μm) penetrate predominantly intothe peripheral region of the lungs.

In one aspect of the invention the treatment methodology begins withparticles of a given size, carries out treatment for a given period oftime after which the particles are increased in size. The particlesinitially administered to the patient penetrate deeply into the lung,i.e., the smallest particles (e.g., 0.5 to 2 microns (μm)) target thealveolar ducts and the alveoli. When the deepest part of the lung istargeted with the smallest particles the patient receives an immediate“rush” from the nicotine delivered which closely matches that receivedwhen smoking a cigarette. These small particles can be obtained by anymethod that produces inhalable particles, such as by milling powder intothe desired size and inhaling the powder or by creating a solution orsuspension and aerosolizing the formulation, e.g. by nebulization or bymoving the solution or suspension through the pores of a membrane. Ineither case, the desired result is to obtain particles which have adiameter in the range of 0.5 μm to about 2 μm. Those skilled in the artwill understand that some of the particles will fall above and below thedesired range. However, if the majority of the particles (50% or more)fall within the desired range then the desired area of the lung will becorrectly targeted.

In practicing the present invention, the patient is allowed torepeatedly administer the tobacco-less formulation of the invention whena cigarette is desired. For example, the patient would be instructed torepeatedly administer the tobacco-less formulation when the patientwould normally smoke a cigarette. In this manner, the patient willbecome accustomed to finding that the device administers nicotine intothe patient in the same manner that a cigarette does. In one embodimentof the invention the concentration of the nicotine in the tobacco-lessformulation could be reduced gradually over time. This could be doneover a sufficiently long period of time so as to allow the patient towean off of nicotine. However, in another embodiment of the inventionthe amount of nicotine is kept substantially constant but the size ofthe aerosolized particles created are increased.

In another treatment methodology, the patient would begin the treatmentwith a low dose of the tobacco-less formulation of the invention andthis dosage would gradually be raised as the patient grew more tolerantof the formulation. With the increasing tobacco-less formulation dosage,the patient could gradually cease smoking until the tobacco-lessformulation completely replaced the cigarette. Administration of aconstant dose of antidepressant or anxiolytic throughout this processmay further improve the patient's probability of overall success. Oncethe cigarette habit is broken, the patient would gradually lower thedosage of the tobacco-less formulation until the nicotine addiction wasbroken. The continued use of the antidepressant or anxiolytic couldenhance the patient's ability to wean themselves off the tobacco-lessnicotine formulation.

Another treatment methodology would gradually increase the size of theparticles for the first form of nicotine. The increased particle sizetargets predominantly the respiratory tract above the alveolar ducts andbelow the small bronchi. This can generally be accomplished by creatingaerosolized particles of nicotine which have a size and range of about 2μm to about 4 μm. Administration is carried out in the same manner asdescribed above. Specifically, the patient administers the aerosolizednicotine at the same time when the patient would be smoking a cigarette.Since the patient has become adjusted to receiving the nicotine “rush”from the smaller sized particles, the patient will expect and istherefore likely to experience the same “rush” when administering theslightly larger particles. However, the effect will be less immediate asa consequence of the particles being deposited predominantly in a higherregion of the respiratory tract. This procedure is carried out over aperiod of time, e.g., days or weeks. In one embodiment of the inventionit is possible to reduce the dose of aerosolized nicotine delivered tothe patient during this second phase. However, the dose may remainconstant.

The treatment can be completed after any phase, e.g. after the secondphase. However, in accordance with a more preferred embodiment of theinvention a third phase of treatment is carried out. Within the thirdphase the particle size of the first form of nicotine is increasedagain. The particles are increased to a size in a range from about 4 μmto about 8 μm or, alternatively, perhaps as large as 12 μm. These largerparticles will target predominantly the upper airways. The largerparticles will give a very small immediate “rush” but will still beabsorbed through the mucous membranes of the patient's respiratorytract. Accordingly, the patient will be administering nicotine doseswhich may be the same as those doses administered at the beginning oftreatment. At this point the treatment can take a number of differentdirections. The patient can attempt to stop administration by immediateand complete cessation of nicotine delivery. Alternatively, the patientcan try to wean off of nicotine by delivering fewer doses during a giventime period, or by decreasing the dose per use, as discussed below.

In another alternative, the same size dose (volume of aerosolformulation) is administered and delivered, creating the same amount ofaerosol, but wherein the aerosolized particles contain progressivelyless nicotine (e.g., more dilute concentration of nicotine in theparticles or dropets). The amount of nicotine can be decreased until thepatient is receiving little or no nicotine. Those skilled in the artreading this disclosure will recognize variations on the overall methodand methods for stopping treatment.

In yet another alternative embodiment the amount of nicotine,concentration of nicotine and particle sizes created by the formulationare all maintained the same from one group of packets to the next.However, the pH of the formulation within the packets from one group tothe next is changed and is generally changed from a high or basic pH toa low or acidic pH. Thus, for example, the pH of the packets within afirst group could be at 9.0 and the pH of the formulation in a secondgroup of packets could be 8.0, followed by a third group at 7.0 followedby a fourth group at 6.0 followed by a fifth group at 5.0. Those skilledin the art, reading this disclosure will understand that the variationin pH from one group to the next can be in any amount and the pH canbegin and end at any point provided the resulting formulation does notcause damage to the lungs of the patient to an unacceptable degree. Inpreferred embodiments, the pH of the first form of nicotine is variedfrom basic to acid thereby gradually decreasing the amount of free basenicotine in the formulation. The pH of the second form of nicotine mayalso be adjusted, but preferably remains constant, typically at aneutral or acidic pH level.

In yet another embodiment of the invention the nicotine forms of theinvention may include variations of all or any of the differentparameters which include amount of nicotine, concentration of nicotine,particle size of aerosol created and pH of the formulation. Any one,two, three or four of the parameters can be varied from oneadministration to the next.

Supplemental Treatment Methodology

Tobacco users wishing to quit may be treated solely with respiratorynicotine as indicated above, i.e. by intrapulmonary delivery. However,it is possible to treat such patients with a combination of pulmonaryadministration and other means of administration, such as transdermaladministration. Transdermal nicotine is preferably administered tomaintain a steady state level of nicotine within the circulatory system.Nasal or buccal formulation could be used for nasal or buccal deliverywhich could supplement aerosolized delivery.

Supplemental nicotine treatments may be combined with administration ofthe formulations of the present invention in an effort to augmentnicotine treatment. As noted above, the exemplary supplementaltreatments identified below typically do not provide the rapid increasein arterial nicotine concentration provided by the formulation of thepresent invention. The supplements do however generally provide a basalsustained nicotine level that contributes to the effect of the secondforms of nicotine of the present invention. The supplemental nicotinetreatments listed below are exemplary only and do not constitute anexhaustive list.

By way of example, transdermal nicotine delivery systems includenicotine patches and others that are described in the art for example,in U.S. Pat. Nos. 4,597,961, 5,004,610, 4,946,853, and 4,920,989, eachof which is expressly incorporated herein by reference.

Transmucosal administration is also known, for example delivery ofnicotine to the systemic circulation through oral drug dosage forms(e.g., lozenge, capsule, gum, tablet, suppository, ointment, gel,pessary, membrane, and powder) are typically held in contact with themucosal membrane and disintegrate and/or dissolve rapidly to allowimmediate systemic absorption. This term includes, but is not limitedto, lozenges, capsules, tablets, and gum. These formulations can also besuch that they provide slow, sustained release of nicotine yieldingprolonged arterial blood concentrations of nicotine.

Preferably, the orally administrable nicotine formulation will consistof any lozenge, tablet, capsule, or gum formulation that deliversnicotine through the oral mucosa cavity, and preferably through thebuccal and/or sublingual mucosa. The nicotine form that is added orincorporated into the nicotine formulations may be pure nicotine or anycompound thereof. The method of manufacture of these formulations may beany suitable method known in the art, including but not limited to theaddition of a nicotine compound to premanufactured tablets; coldcompression of an inert filler, a binder, and either pure nicotine or anicotine-containing substance (as described in U.S. Pat. No. 4,806,356,herein incorporated by reference); encapsulation of nicotine or anicotine compound; and incorporation of nicotine bound to a cationexchange resin, for example, as in a chewing gum (as described in U.S.Pat. Nos. 3,877,468 and 3,901,248, herein incorporated by reference).

Based on the above, it will be understood by those skilled in the artthat a plurality of different treatments and means of administration canbe used to treat a single patient. For example, a patient can besimultaneously treated with nicotine by transdermal administration,nicotine via pulmonary administration, in accordance with the presentinvention, and nicotine which is administered to the mucosa.

IV. Nicotine Delivery Devices

The aspects of the invention described above such as changing theamount, concentration, or pH of the formulation or changing the particlesize of the aerosol created with the formulation can be done independentof the delivery device. However, there are a number of features whichcan be included in the system which are specific to the device whichdelivers the formulation. For example, the device can be designed so asto avoid overdosing. This can be carried out by electronicallymonitoring the number of doses a patient has delivered and locking outfurther use for a given time interval. Thus, this system can be used asa safety feature. In addition to a safety feature the device can beprogrammed in order to force the frequency of administration. This couldbe done in order to aid the patient in reducing the times the dose isdelivered and thereby moving the patient forward towards a point in timewhen the patient no longer needs nicotine.

Devices, if desired, contain a variety of components to facilitate thedelivery of the formulations of the invention. For instance, the devicemay include any component known in the art to control the timing of drugaerosolization relative to inhalation (e.g., breath-actuation), toprovide feedback to patients on the rate and/or volume of inhalation, toprevent excessive use (i.e., “lock-out” feature), to prevent use byunauthorized individuals, and/or to record dosing histories.

Any of the devices suitable for use with the invention could be designedto force the patient to use only a certain dosage form of thetobacco-less formulation for a given period of time and then requirethat the patient use another dosage form. In this way the device can beprogrammed to start the patient with, for example, a relatively highdose which can be quickly administered and thereafter allowing thedevice only to be activated when a second group with a smaller amount,lower concentration, etc. is used in the device.

The devices suitable for use with the invention can also be programmedto be patient and physician specific. Thus, the device can include alock-out component which prevents the device being used except in thepresence of another component which could, for example, be a wristbandworn by the patient. The device could also be programmable only by aparticular physician equipped with a device which sends a signalallowing the device to be reprogrammed.

Devices suitable for use with the invention can also be programmed torelease larger or lesser amounts of formulation and fire the aerosol atdifferent rates. Either or both of these parameters can be changed bythemselves, together or in combination with the other parametersrelating to the formulation and particle size.

Although any device suitable for delivering the requisite amounts offormulation to the lungs of a patient may be employed to deliver theformulations of the invention, the Aradigm AERx Essence® is preferred.

Precision delivery of small molecule drugs via the lung for systemiceffect is possible. An electronic inhaler capable of delivering a liquidformulated drug stored in a unit dose packages has been described anddisclosed in U.S. Pat. No. 5,718,222 entitled “Disposable Package forUse in Aerosolized Delivery of Drugs,” and is incorporated herein byreference. A formulation of nicotine can be prepared for delivery withthis system. Quantitative delivery of nicotine on demand provides amechanism for nicotine replacement therapy which is unlikely to beassociated with recidivism precipitated by the symptoms of physicalwithdrawal.

In one embodiment, the tobacco-less nicotine formulation of theinvention is forced through the openings or pores of a porous membraneto create an aerosol. In a specific embodiment, the openings are alluniform in size and are positioned at uniform distances from each other.However, the openings can be varied in size and randomly placed on themembrane. If the size of the openings is varied, the size of theparticles formed will also vary. In general, it is preferable to havethe opening sizes within the range of about 0.25 μm to about 6 μm whichwill create particle sizes of about 0.5 μm to 12 μm which are preferredwith respect to inhalation applications. When the openings have a poresize in the range of 0.25 μm to 1 μm they will produce an aerosol havingparticle sizes in the range of 0.5 μm to 2 μm, which is particularlyuseful for delivering nicotine to the alveolar ducts and alveoli. Poresizes having a diameter of about 1 μm to 2 μm will produce particleshaving a diameter of about 2 μm to 4 μm, which are particularly usefulfor delivering nicotine to the area above the alveolar ducts and belowthe small bronchi. A pore size of 2 μm to 4 μm will create particleshaving a diameter of 4 μm to 8 μm, which will target the area of therespiratory tract from the small bronchi upward.

Increasing the size of the openings of the porous membranes producesnicotine formulation particles of increasing size. A strategy in whichthe blood levels of nicotine, and especially the peak levels, arereduced gradually will be the most effective in treating the symptoms ofwithdrawal, and thereby increase the chances of successful smokingcessation. In one embodiment of the invention, the size of theaerosolized nicotine particles is increased in a stepwise manner byusing porous membranes that create “monodisperse” aerosols, wherein allthe particles within the aerosol created have essentially the sameparticle size. Nicotine particles of increasing size are produced byusing membranes of increasing pore sizes.

In another embodiment, the size of aerosolized tobacco-less nicotineformulation particles is increased in gradient fashion by using porousmembranes that create “multi-disperse” aerosols, wherein the particleswithin the aerosol created have different particle sizes. Membraneswhich have an increasing range of pore sizes are used to producenicotine particles of increasing size.

As intrapulmonary administration is not 100% efficient, the amount ofdrug aerosolized will be greater than the amount that actually reachesthe patient's circulation. For example, if the inhalation system used isonly 50% efficient then the patient will aerosolize a dose which istwice that needed to raise the patient's nicotine level to the extentneeded to obtain the desired results. More specifically, when attemptingto administer 1 mg of nicotine with a delivery system known to be 50%efficient, the patient will aerosolize an amount of formulationcontaining about 2 mg of nicotine.

A device comprised of a container that includes an opening covered by aporous membrane, such as the device disclosed in U.S. Pat. No.5,906,202, may be used to deliver nicotine. The device may be designedto have the shape and/or bear the markings of a pack of cigarettes, andmay include the scent of tobacco. These features and others that addressthe behavioral component of cigarette smoking may enhance theeffectiveness of the method described herein.

Containers for the formulations of the present invention may be any formsuitable for use with the chosen delivery system. Preferred containersfor formulations designed to be delivered as aerosols are single dosepackets, for example blister packets containing a liquid sterileformulation of the invention. In one embodiment, the volume of thereceptacle is at least about 0.037 cm³. In another embodiment, thevolume of the receptacle is at least about 0.048 cm³. In yet anotherembodiment, are receptacles having a volume of at least about 0.067 cm³or 0.095 cm³. In one embodiment of the invention, the receptacle is acapsule that holds try powder containing nicotine designated with acapsule size 2, 1, 0, 00 or 000. Suitable capsules can be obtained, forexample, from Shionogi (Rockville, Md.). Blisters designed to holdpowder formulations can be obtained, for example, from Hueck Foils,(Wall, N.J.).

V. Dosing

A tobacco cigarette contains 6 to 11 mg of nicotine, of which the smokertypically absorbs 1 to 3 mg; see Henningfield N Engl J Med 333:1196-1203(1995). Factors influencing nicotine absorption includesubject-dependent factors, such as smoking behavior, lung clearancerate, etc., morphological factors, and physiological factors, such astidal volume, inspiratory and expiratory flow rate, particle size anddensity. See Darby et al., Clin Pharmacokinet 9:435-439 (1984). Thesystemic dose of nicotine per puff is extremely variable, however, peakplasma concentrations of 25 to 40 ng/mL of nicotine, achieved within 5to 7 minutes by cigarette smoking, are believed typical. In accordancewith the present invention, about 0.05 to about 3 mg, preferably about0.3 to about 1 mg, preferentially about 0.3 to about 0.7 mg of nicotineare delivered to the lungs of the patient in a single dose to achievepeak blood plasma concentrations of 10 to 40 ng/mL. These specificamounts should not be relied on. Alternatively, the amounts should bemeasured, adjusted, remeasured and readjusted as needed to obtain theappropriate dosing. An aspect of the invention is to initially set outto deliver the nicotine preparation in a manner that satisfies thecraving for high plasma levels of nicotine in the subject and thengradually changing the nature of the inhaled nicotine formulation interms of the amount of nicotine, its concentration as well as site ofdeposition so as to gradually reduce the peak plasma nicotine levels towean the subject off tobacco smoking. The amount needed will vary basedon many factors including how much the patient smokes, and the patient'sage, sex, weight and condition.

The amount of nicotine administered will vary based on factors such asthe age, weight and frequency of smoking or nicotine tolerance of thesmoker. Other factors, such as daily stress patterns, and demographicfactors may also help to determine the amount of nicotine sufficient tosatisfy the smoker's craving for the drug. Administering nicotine usingthe methods of the present invention can involve the dailyadministration of anywhere from 0.05 mg to 200 mg of nicotine, but morepreferably involves the administration of approximately 1 to 100 mg perday, but these amount ranges should not be relied on. Amounts should bedetermined as indicated above.

When nicotine enters the circulatory system of a human patient it isoxidized to cotinine within four to six hours. The present inventionincludes the administration of cotinine and other nicotine derivativesprovided such derivatives do not result in unacceptable adverse effects.

Methods of Administering Formulations

The tobacco-less formulations described herein may be administered bysystemic injection, transdermal administration by applying themedicament directly to the skin, oral ingestion, inhalation as describedherein, or by other methods such as systemic infusion. Commerciallyavailable nebulizers for liquid formulations, including jet nebulizersand ultrasonic nebulizers may be useful for administration. Liquidformulations may be directly nebulized and lyophilized power nebulizedafter reconstitution. Alternatively the tobacco-less formulation may beaerosolized using a metered dose inhaler, or inhaled as a powder thatcould be prepared by any of the methods known to those skilled in theart. For example, the powder could be prepared by lyophilization,spray-drying, freeze-drying, milling, or by incorporation of nicotineinto premanufactured particles or entrapment in microparticles. Inaddition, a liquid medicament may be directly instilled in thenasotracheal or endotracheal tubes in intubated patients.

Effective dosages and schedules for administering the formulations maybe determined empirically, and making such determinations is within theskill in the art Those skilled in the art will understand that thedosage of tobacco-less formulation of the invention that must beadministered will vary depending on, for example, the person receivingthe formulation, the route of administration, the particular type offormulation used and other drugs being administered to the patient. Aspreviously noted, the formulation of the present invention may beadministered in a single dose, or as multiple doses over time.

The formulations are typically administered in a dose sufficient toprovide a therapeutically effective level. By way of example, nicotinearterial concentration produced by the first nicotine form is preferablyat least 10 ng/ml, but may be 15, 20, 25, 30, 35, 40, 50 or more ng/mlnicotine, being limited by the amount necessary to address the nicotineaddiction while not reaching toxic levels. Similarly, the second form ofnicotine is administered in an amount to maintain a second form ofnicotine arterial concentration in the patient for at least 60 minutesafter administration. This may be augmented by addition of slow releasecomponents such as cyclodextrin, encapsulation of the active nicotine,chemical or physical modification of the form of nicotine and the likeas described herein. The maintained arterial concentration of the secondform of nicotine is at least 5 ng/ml, preferably 7, 10, 12, 15 or 20ng/ml.

It would be apparent to a person skilled in the art that variations maybe acceptable with respect to the therapeutically effective dose andfrequency of the administration of formulations of the invention. Forexample, the amount of the formulation administered may be inverselycorrelated with the frequency of administration. For example, anincrease in the concentration of neurologic agent in a singleadministered dose, or an increase in the mean residence time in the caseof a sustained release form of neurologic agent, generally will becoupled with a decrease in the frequency of administration.

It is appreciated by those of skill in the art that the actualformulation dose will depend on a variety of factors that may bespecific to the subject undergoing dosing. These factors should be takeninto consideration when determining the therapeutically effectiveformulation dose and frequency of its administration. For example, theeffective dose can depend on the age, weight, or general health of thesubject; the severity of the nicotine addiction; the frequency andduration of dosing; the type of formulation administered; thecharacteristics, such as lipophilicity, of the formulation andcomposition; and the like. Generally, a higher dosage is preferred ifthe nicotine addiction is more severe. Thus some minor degree ofexperimentation may be required to determine the most effective dose andfrequency of dose administration, this being well within the capabilityof one skilled in the art once apprised of the present disclosure.

Intermittent Dosing

In another embodiment of the invention, the therapeutically effectiveformulation is administered intermittently. “Intermittentadministration” is intended administration of a therapeuticallyeffective formulation dose followed by a time period of discontinuance,which is then followed by another administration of a therapeuticallyeffective dose, and so forth. “Time period of discontinuance” isintended a discontinuing of daily administration of the formulation.During the time period of discontinuance, the arterial nicotine plasmaconcentration is substantially below the maximum level obtained duringtreatment. The preferred length of the discontinuance period depends onthe concentration of the effective formulation dosage and the form ofthe formulation used. The discontinuance period can be at least 2 days,preferably is at least 4 days, more preferably is at least 1 week andgenerally does not exceed a period of 4 weeks unless the patient hasovercome the addiction to nicotine. An intermittent schedule ofadministration of agent can continue until the desired therapeuticeffect, and ultimately treatment of the addition, is achieved.

In yet another embodiment, intermittent administration of thetherapeutically effective formulation dose is cyclic. By “cyclic” isintended intermittent administration accompanied by breaks in theadministration, with cycles ranging from about 1 week to about 2, 3, 4,5, or 6 weeks, more preferably about 2 weeks to about 4 weeks. Forexample, the administration schedule might be intermittentadministration of the effective formulation dose with a single dose isgiven three times per week for 4 weeks, followed by a break inintermittent administration for a period of a week, followed byintermittent administration by administration of a single dose givenonce per week for 3 weeks, and so forth. A cyclic intermittent scheduleof administration of the formulation to a patient may continue until thenicotine addiction is overcome.

VI. Assessing Addiction

A variety of methods may be utilized to assess the craving for nicotine,including but not limited to, the nicotine craving test specified by theDiagnostic and Statistical Manual of Mental Disorders, Revised ThirdEdition (DSM-III-R) (see (1991) J. Am. Med. Assoc. 266:3133); theShiffman-Jarvik Craving Subscale (see O'Connell and Martin (1987) J.Consult. Clin. Psychol. 55:367-371 and Steur and Wewers (1989) ONF16:193-198, also describing a parallel visual analog test); West et al.(1984) Br. J. Addiction 79:215-219; and Hughes et al. (1984)Psychopharmacology 83:82-87, each of which is expressly incorporatedherein by reference.

A preferred nicotine craving scale is that specified in DSM-III-R,supra. According to this scale, a subject is asked to rate the severityof his craving for nicotine on a scale between 0 and 4, wherein 0 isnone; 1 is slight; 2 is mild; 3 is moderate; and 4 is severe. Using thecompositions and methods described herein, the subject should attain atleast a one unit, and preferably at least a two unit, decrease in hiscraving for nicotine as measured by the protocol set forth in DSM-III-Rfrom about 2 to 30 minutes after administration of the oral nicotineformulation. More preferably, the maximum reduction in craving fornicotine will occur from about 2 to 20 minutes, and more preferably fromabout 2 to 10 minutes after administration of the oral nicotineformulation.

The Shiffman-Jarvik Craving Scale is a six-item, forced-choice,self-report tool that measures cigarette craving. Each item has sevenpossible responses which correspond to scores ranging from 1 (nocraving) to 7 (high craving). A mean score is obtained to determine therespondent's level of craving. A typical craving score measured 48 hoursafter the initiation of a smoking cessation program is between about 4and 5; while a two-week follow-up craving scale will typically bebetween about 3 and 4. Using the compositions and methods describedherein, the subject should attain at least a one unit, and preferably atleast a two unit, decrease in his craving for nicotine as measured bythe protocol set forth in the Shiffman-Jarvik Craving Scale from about 2to 30 minutes after administration of the oral nicotine formulation.More preferably, the maximum reduction in craving for nicotine willoccur from about 2 to 20 minutes, and more preferably from about 2 to 10minutes after administration of the oral nicotine formulation.

The “craving questionnaire” craving scale employs a five itemquestionnaire that asks subjects to rate how much they had been missingtheir cigarettes, how difficult it had been to be without cigarettes,how much they had been aware of not smoking, how pre-occupied they hadbeen with thinking about cigarettes, and how much they had craved theircigarettes. The subject responds to each question with a number between1 and 3, where 1 is low and 3 is high. The ratings are combined to givea single craving score. According to this craving scale, a combinedscore of between about 9 and 12 is typical. Using the compositions andmethods described herein, the subject should attain at least a threeunit, and preferably at least a four unit, decrease in his craving fornicotine as measured by the protocol set forth for use with this cravingquestionnaire from about 2 to 30 minutes after administration of theoral nicotine formulation. More preferably, the maximum reduction incraving for nicotine will occur from about 2 to 20 minutes, and morepreferably from about 2 to 10 minutes after administration of the oralnicotine formulation.

A subject's nicotine dependence can be quantified using aneight-question scale, termed the Fagerstrom Nicotine Tolerance Scale(see Fagerstrom (1978) Addict. Behav. 3:235-241 and Sachs (1986) Clinicsin Geriatric Medicine 2:337-362) which provides a relative index of thedegree of physical dependency that a patient has for nicotine. This testis shown in FIG. 4.

These tests have a variety of uses in practicing the instant invention.For example, the Fagerstrom test may be used to estimate nicotinetolerance and therefore the initial nicotine dose in treatment. Cravingsscores may be used to determine the effectiveness of a given formulationdosage in suppressing the desire to smoke or chew tobacco.

As will be evident to one of skill in the art, the ability to measurethe patient's arterial nicotine plasma levels can be of tremendous valuein tailoring a smoking cessation or other therapy to the patient'sneeds. There has been very little discussion in the literature of usingdirect or indirect measurement of arterial nicotine levels as anintegral part of smoking cessation therapy. The traditional interest inquantifying arterial nicotine levels has been related to research onefficacy of smoking cessation therapies. For example, research studiescommonly used various measurement techniques to attempt to verifyself-reports of smoking frequencies by study subjects. These include themeasurement in saliva and blood plasma of nicotine, cotinine (theprimary metabolite of nicotine), carboxyhemoglobin, and thiocyanate; andthe measurement in expired air of carbon monoxide. The most frequentlycited technique is the quantification of cotinine, a nicotinemetabolite, in saliva. The quantification of cotinine in blood fluidscan be accomplished by gas-liquid chromatography, radioimmunoassay, andliquid chromatography. (For a discussion of liquid chromatographicassays for cotinine, see Machacek and Jiang (1986) Clin. Chem.32:979-982, herein incorporated by references.)

The present invention may optionally include the direct or indirectmeasurement of nicotine blood levels as an integral part of methods fortreating conditions responsive to nicotine therapy, and particularly forsmoking cessation therapy and for reducing nicotine craving. Thenicotine blood levels can be measured before, during, or after theadministration of the formulations of the invention, as an aid indetermining the amount of nicotine to be administered and the frequencyof administration. In a preferred embodiment, saliva samples are takenfrom the patients and used for measurement of cotinine, as a biochemicalmarker of nicotine blood plasma levels. Cotinine levels are determinedusing any of the analytical methods known to those skilled in the art.In a particularly preferred embodiment, the cotinine assay would beportable and easily and simply accomplished by the patient, as in anassay kit or strip indicator.

All publications and patent applications cited in this specification areherein incorporated by reference as if each individual publication orpatent application were specifically and individually indicated to beincorporated by reference.

Although the foregoing invention has been described in some detail byway of illustration and example for clarity and understanding, it willbe readily apparent to one of ordinary skill in the art in light of theteachings of this invention that certain changes and modifications maybe made thereto without departing from the spirit and scope of theappended claims.

As can be appreciated from the disclosure provided above, the presentinvention has a wide variety of applications. Accordingly, the followingexamples are offered for illustration purposes and are not intended tobe construed as a limitation on the invention in any way. Those of skillin the art will readily recognize a variety of noncritical parametersthat could be changed or modified to yield essentially similar results.

EXAMPLES Example 1 Single-Dose Application of Deep Lung NicotineFormulation

Smoking dependence appears partly related to the “high & fast” rise inplasma nicotine concentration achieved by cigarettes. However, unlikecigarettes, current nicotine replacement therapies (NRTs) attainrelatively “low & slow” nicotine plasma levels (FIG. 1). This exampleillustrates that a nicotine delivery system that provides cigarette-likeplasma levels, serves to reduce acute craving, inhibit relapse, andresult in higher smoking cessation rates compared with existing NRTs.

The AERx Essence System known in the art was used to deliversingle-bolus doses of aerosolized nicotine to healthy adult malesmokers. The AERx Essence is an all-mechanical, nonpropellent driven,hand-held device that uses individually packaged, single-use, dosageform strips. A uniformly fine, respirable aerosol is created when thedrug solution is “extruded” through an array of submicron sized holesdrilled into the dosage form strip. The fine aerosol that is generatedallows the deep-lung deposition needed to achieve rapid and efficientabsorption of drug similar to that obtained by smoking. This inhalationdelivery system is a part of the AERx inhalation delivery platforms;AERx devices may be all-mechanical, or electro-mechanical. They may alsohave various electronic components. Some of these devices includediagnostic and disease management tools as well. The AERx Essence deviceactuates the piston movement when the patient pushes a button that alsocauses opening of the valve through which the air that the patient isinhaling enters the device. The inspiratory flow rate is mechanicallycontrolled in this particular embodiment of the AERx Essence device.

Methods

Eighteen healthy, adult male smokers were enrolled in a randomized,open-label, multiple-exposure study which was conducted in two parts.Two subjects were removed prior to Study Part 2 with sixteen subjectsstarting and completing Study Part 2. Subjects' ages ranged from 19-41years (mean=27 years).

In Study Part 1, the tolerability and safety of seven nicotineconcentrations were evaluated. In Study Part 2, subjects received one ofthree nicotine concentrations: 10, 20, or 30 mg/ml, delivering bolusnicotine lung doses of approximately 0.2, 0.4 and 0.7 mg, respectively.Measures of arterial nicotine plasma concentration and acute post-dosingcigarette craving scores (11-point VAS) were made following a singleinhalation of nicotine.

Results

Safety and Tolerability: No clinically significant changes in safetymeasures were noted following dosing (vital signs, ECG, spirometry,labs). A total of 119 adverse events (AEs) were recorded. Most AEs werereported as either mild or moderate and self-resolved withoutmedication. No serious AEs were observed. The most commonly reported AEswere throat irritation, lightheadedness (Table 1).

TABLE 1 Incidence of most common Adverse Events (AE) Adverse Event (AE)Incidence Subjects Experiencing AE Throat irritation 46 17Lightheadedness 22 11 Cough 20 10

Pharmacokinetics: Arterial plasma nicotine pharmacokinetics demonstratedrapid onset (Tmax=1 min) and substantial peak plasma concentrations.Maximum plasma concentrations (Cmax) and area under theconcentration-time curves (AUC) were consistent with a trend toward doseproportionality (FIG. 2, Table 2).

TABLE 2 Mean Nicotine Pharmacokinetic Parameters Parameter 10 mg/ml 20mg/ml 30 mg/ml Tmax (min) 1 1 1 Cmax (ng/ml) 11.5 (9.5) 18.0 (3.6) 22.9(9.0)  T½ (min⁻¹) 136 (58) 114 (18) 97 (16) AUC_(0-t) (ng · min/ml)  319(219)  532 (116) 622 (218) Standard deviations are in parenthesis.

Acute Craving: Patients were asked to rate their nicotine craving on ascale of 0 to 10 pre- and post-dosing. Nearly all subjects reported anacute reduction in craving or an absence of craving immediatelyfollowing study dosing. A mean reduction in craving from baseline wasobserved following all three dose levels (FIG. 3). Combining all doselevels, mean craving declined from 4.9 to 1.4 within 5 minutespost-dosing, and remained below pre-dose baseline for the 4 hours ofmonitoring.

Conclusions

Inhaled nicotine via the AERx Essence appears safe and tolerable. TheAERx Essence delivers inhaled nicotine with a PK profile that isconsistent with the rapid delivery and absorption seen with cigarettesmoking, and acute craving following inhaled nicotine via the AERxEssence appears to be acutely reduced

Example 2 Use of Alternative Nicotine Forms

This example demonstrates the effectiveness of different nicotine dosageforms of the invention. The aim of the example is to illustrate thatgenerically available nicotine formulations are suitable for use in thepresent invention.

Formulation studies were performed to evaluate the effectiveness ofnicotine salts and pH on the stability of nicotine in AERx® dosageforms. Nicotine is a weak base (pKa₁=3.4 and pKa₂=8.4) and in theun-ionized state had the capability to get absorbed into the polymericmaterials used in many nicotine delivery systems. When a screening studywas conducted in the pH 3.0-7.0 range using buffered nicotine sulphateand bitartrate, nicotine concentration was in effect unaltered for thetwo salts at the lower pH's of 3.0 and 4.0. Nicotine bitartrate wasbetter in this pH range as compared to nicotine sulphate in terms ofensuring that there was no loss of nicotine into the polymeric dosageform materials. A theoretical calculation using the Henderson-Hasselbachequation indicated that the ratio of ionized to un-ionized species at pH3.0 and pH 4.0 was 158489 and 15849, respectively, implying limitedpotential for absorption to occur at the lower pH of 3.0.

Aradigm's proprietary AERx® System was used in the present example. Thissystem consists of the AERx® Strip™, a single-use disposable dosageform, and the AERx® device, which has two hand-held configurations: anelectromechanical version and an all-mechanical version.

Nicotine formulations were packaged under aseptic conditions into theAERx® Strip, to create a sterile dosage form. Aerosol generation usingthe AERx® System is completed in one or two seconds via mechanicalpressurization of the nicotine formulation. This pressurization causesthe seal in the AERx® Strip between the drug reservoir and a nozzlearray to peel open. This leads to the nicotine formulation beingexpelled through the nozzle array as a fine aerosol. By varying the sizeof the nozzle holes, the size of the aerosol can be modified to optimizeregional lung deposition. The electromechanical AERx® system wasmodified to allow addition of dose titration capabilities into thesystem for this program.

Results

Analytical Assay Development for Nicotine Quantitation

A high performance liquid chromatography (HPLC)-based assay wasdeveloped in house to enable quantitation of nicotine (Table 3). TheHPLC method was suitably modified for functional (aerosol) testing ofAERx®-nicotine and a partial qualification conducted. The analyticalperformance parameters evaluated were: standard linearity, range,accuracy, precision, limit of quantitation (LOQ), system suitability,specificity and solution stability. The functional test method, inconjunction with the RP-HPLC method was qualified for use in determiningemitted dose and particle size distribution of aerosolized nicotine.Nicotine working standard linearity, r2, was 1.000 and the linearconcentration range was 0.5 to 40.0 μg/mL (Table 4).

TABLE 3 Analytical Method Parameters/Details Reverse Phase HighPerformance Liquid Chromatography (RP-HPLC) Method HPLC Column Ace 5,C18 (25 cm × 4.6 mm, 5 μm) Mobile Phase 80% 20 mM Phosphate buffer, 20%Methanol, pH 5.0 Wavelength (UV 259 nm detector) Flow Rate 1.00 mL/minInjection Volume 20 μL Column Temperature 35° C. Autosampler AmbientTemperature Run Time 10 minutes

TABLE 4 Summary of analytical results from method development AnalyticalPerformance Parameters Evaluated Results Standard Linearity and Range R²= 1.000, 0.5-40.0 μgmL Accuracy and Precision Passed acceptance criteriaLimit of Quantitation 0.5 μg/mL System Suitability and Peak Area & RT: %RSD < 2%, Tailing Specificity Factor = 1.0, No interfering peak SolutionStability Standards Stability = 7 days, Diluent/Mobile Phase Stability =15 days

Nicotine Formulation Development

Selecting Nicotine salts: After evaluation of availability of variousgrades of nicotine salts on the market, nicotine bitartrate and nicotinesulphate were selected for further screening. Both salts were purchasedfrom Nicobrand Limited, Northern Ireland.

Formulation Concentrations: A 0.9-1.0 mg lung dose was estimated as anefficacious upper end dose based on available literature. Estimating a60% deep lung delivery efficiency for AERx®, the nicotine concentrationchosen at the upper end was 32.0 mg/mL. Using the three step dosereduction strategy described above, the lower nicotine concentration wasestimated to be 10.7 mg/mL. Initial formulation studies used a lowerconcentration of 8.0 mg/mL (prior to the finalization of a three-stepdose reduction strategy), which was later finalized (using a three stepdose reduction strategy) to be 10.7 mg/mL.

Formulation stability in pouches: An initial formulation screening studywas initiated utilizing nicotine formulations between the pH of 3.0-7.0stored in pouches at 40° C./75% R.H. The pouches were made of the samepolymeric material as the contact layer in AERx® dosage forms. Inprevious studies with a different but chemically similar drug, polymericmaterials showed the potential for absorptive losses of drug fromsolution. Nicotine concentration as well as pH was monitored for aperiod of 28 days.

Results indicated no impact on pH over the 28 days period throughout thepH range evaluated (Tables 5 & 6). The concentration of nicotinedecreased over time at higher pH values, consistent with the proposedabsorption when in the unionized form (Tables 7 & 8). The concentrationof nicotine was unaltered at pH's 3.0 and 4.0.

TABLE 5 Formulation stability- nicotine bitartrate pH results in pouchesNicotine Bitartrate (controls) Nicotine Bitartrate (pouches) 8 mg/mL 32mg/mL 8 mg/mL 32 mg/mL T = 7 T = 15 T = 28 T = 7 T = 15 T = 28 T = 7 T =15 T = 28 T = 7 T = 15 T = 28 Theoretical pH T = 0 days days days T = 0days days days T = 0 days days days T = 0 days days days 3.0 3.0 3.1 3.13.1 3.0 3.1 3.1 3.1 3.1 3.1 3.3 3.2 3.1 3.1 3.2 3.2 4.0 4.0 4.3 4.2 4.24.0 4.3 4.3 4.3 4.3 4.2 4.3 4.4 4.3 4.2 4.3 4.4 5.0 5.1 5.0 5.3 5.3 5.05.3 5.3 5.3 5.4 5.3 5.2 5.2 5.4 5.3 5.2 5.2 6.0 6.0 6.3 6.1 6.2 6.0 6.36.2 6.2 6.3 6.2 6.1 6.1 6.3 6.2 6.0 6.1 7.0 7.1 7.1 7.4 7.3 7.0 7.1 7.27.2 7.4 7.1 7.1 7.1 7.2 6.9 7.0 6.8

TABLE 6 Formulation stability- nicotine sulphate pH results in pouchesNicotine Sulphate (controls) Nicotine Sulphate (pouches) 8 mg/mL 32mg/mL 8 mg/mL 32 mg/mL T = 7 T = 15 T = 28 T = 7 T = 15 T = 28 T = 7 T =15 T = 28 T = 7 T = 15 T = 28 Theoretical pH T = 0 days days days T = 0days days days T = 0 days days days T = 0 days days days 3.0 3.0 2.7 2.82.8 3.0 2.7 2.8 2.8 2.9 2.7 2.8 2.8 2.8 2.8 2.9 2.8 4.0 4.0 4.1 4.1 4.14.1 4.0 4.0 4.1 4.1 4.0 4.1 4.1 4.0 4.0 4.1 4.1 5.0 5.0 5.1 5.1 5.1 5.15.1 5.1 5.1 5.2 5.1 5.0 5.1 5.2 5.1 5.0 5.0 6.0 6.0 6.2 6.1 6.1 6.0 6.26.1 6.1 6.2 6.1 6.0 6.1 6.2 6.0 5.9 6.0 7.0 7.0 7.3 7.1 7.1 7.0 7.2 7.17.1 7.1 6.9 6.9 6.9 7.1 6.9 6.9 6.8

TABLE 7 Formulation stability- nicotine bitartrate concentration resultsin pouches Nicotine Bitartrate Formulation % Recovery (8 mg/mL) %Recovery (32 mg/mL) Theoretical pH T = 0 T = 7 days T = 15 days T = 28days T = 0 T = 7 days T = 15 days T = 28 days 3.0 101.4 99.9 100.0 101.6101.7 99.1 101.9 102.3 4.0 100.8 99 99.0 100.2 100.5 101.9 99.1 98.7 5.0100.6 96.8 97.1 98.3 100.5 99.2 95.4 98.9 6.0 101.7 93.2 90.1 95.8 99.293.1 94.7 95.3 7.0 97.6 72.3 73.5 75.0 98.6 85.8 89.3 87.3

TABLE 8 Formulation stability- nicotine sulphate concentration resultsin pouches Nicotine Sulphate Formulation % Recovery (8 mg/mL) % Recovery(32 mg/mL) Theoretical pH T = 0 T = 7 days T = 15 days T = 28 days T = 0T = 7 days T = 15 days T = 28 days 3.0 100.3 94.9 96.9 102.4 98.1 101.2100.2 99.0 4.0 99.5 95.4 98.3 100.9 98.4 97.0 98.3 99.0 5.0 100.4 95.695.2 98.5 98.5 97.8 99.0 98.0 6.0 100.2 94.3 92.5 94.0 99.3 94.6 95.095.8 7.0 97.6 75.0 75.2 82.2 99.0 89.6 89.4 90.1

Based on these results as well as theoretical calculations, pH 3.0 waschosen for use with polymeric products as the proportion of ionizedspecies is maximized at this pH while maintaining acceptable safetyprofiles for an inhaled product.

Formulation stability/screening in AERx® dosage forms: As buffering atextreme pH's is not desirable for inhaled products because it can elicithyperreactivity, pH adjustment is preferred. For this reason anunbuffered formulation was evaluated.

AERx® dosage forms were filled with nicotine bitartrate and nicotinesulphate at both 10.7 and 32.0 mg/mL of nicotine and stored at 40°C./15% R.H. (accelerated storage condition recommended forsemi-permeable containers, ICH Q1A) for a period of 14 days.

The results for pH (Table 9) and concentration (Table 10) indicatedexcellent control, confirming the choice of an unbuffered formulation.Having developed a robust formulation, we then proceeded to evaluate thedose titration capabilities as well as optimizing aerosol performanceusing these formulations.

TABLE 9 Nicotine in AERx ® strips (stored at 40° C./15% RH) pH valuesFormulation T = Initial T = 7 days T = 14 days Nicotine Bitartrate 3.02.9 2.9 (10.7 mg/mL, pH 3.0) Nicotine Bitartrate 3.0 3.0 2.9 (32.0mg/mL, pH 3.0) Nicotine Sulphate 3.0 2.9 2.9 (10.7 mg/mL, pH 3.0)Nicotine Sulphate 3.0 2.9 2.9 (32.0 mg/mL, pH 3.0)

TABLE 10 Recovery of nicotine in AERx ® strips stored at 40° C./15% RH %Recovery (SD) Formulation T = Initial T = 7 days T = 14 days NicotineBitartrate  98.9 (0.5) 102.1 (0.3)  99.3 (0.2) (10.7 mg/mL, pH 3.0)Nicotine Bitartrate 100.2 (0.8) 100.2 (0.5) 102.9 (6.5) (32.0 mg/mL, pH3.0) Nicotine Sulphate 100.0 (0.4) 100.5 (0.2) 100.7 (1.1) (10.7 mg/mL,pH 3.0) Nicotine Sulphate  97.4 (2.3) 100.6 (0.9) 100.0 (0.7) (32.0mg/mL, pH 3.0)

Optimization of aerosol performance of nicotine formulation with theAERx® System

Characterization and optimization of delivery efficiency (emitted dose)of nicotine formulations from AERx® in a simulated inhalation:

Efficiency of delivery of formulation from the AERx® System is expressedas emitted dose (ED). For ED quantification, a known dose of eachnicotine formulation was loaded into AERx® Strips and then aerosolizedonto standardized collection filters. The filters were rinsed thoroughlywith the assay diluent. Spiking studies were conducted to verify thatall of the nicotine was recovered from the filter. The amount ofnicotine in the rinsate was quantified by HPLC.

The ED data was excellent for the partial extrusion as well as multipleconcentrations dose reduction strategies evaluated. Emitted dose inpercent at the three levels using the partial extrusion strategy was20.4, 17.2 and 18.8 with standard deviations of 1.4, 0.8 and 1.0respectively (see Table 11). The percent emitted dose for the successiveconcentrations of 32.0, 21.3 and 10.7 mg/mL was 60.0, 61.7 and 62.7 withthe standard deviations being 3.0, 2.8 and 3.2 respectively (see Table12).

TABLE 11 Emitted dose performance using partial dose settings using 32mg/mL nicotine bitartrate Level 1 Level 2 Level 3 % 2nd % 3rd DF# ED (%LC) ED (% LC) ED (% LC) Total ED % 1st shot shot shot  1 18.7 14.7 18.752.2 35.8 28.3 35.9  2 20.8 17.1 20.2 58.1 35.8 29.4 34.8  3 18.3 17.218.8 54.2 33.7 31.7 34.6  4 19.9 17.1 18.8 55.8 35.7 30.6 33.7  5 21.217.2 20.0 58.4 36.3 29.4 34.2  6 20.6 18.2 19.3 58.1 35.4 31.4 33.2  719.9 17.7 18.3 55.8 35.6 31.6 32.8  8 18.3 17.5 18.3 54.2 33.8 32.3 33.9 9 22.8 17.4 19.5 59.8 38.2 29.1 32.6 10 20.6 17.6 19.4 57.7 35.7 30.633.7 11 20.3 17.0 19.1 56.4 36.1 30.1 33.8 12 22.0 17.1 20.5 59.6 37.028.6 34.4 13 20.6 17.4 17.8 55.8 36.9 31.2 31.9 14 21.0 17.1 17.7 55.837.6 30.7 31.7 15 20.4 17.7 18.8 56.9 35.8 31.1 33.1 16 20.3 16.9 17.354.5 37.3 31.0 31.7 17 22.2 17.3 17.9 57.4 38.6 30.1 31.2 18 21.6 18.318.7 58.7 36.9 31.2 31.9 19 20.2 17.8 20.5 58.4 34.6 30.4 35.0 20 17.515.4 16.8 49.7 35.2 31.0 33.7 Mean 20.4 17.2 18.8 56.4 36.1 30.5 33.4 SD1.36 0.82 1.03 2.53 1.29 1.07 1.27

TABLE 12 Emitted dose performance of nicotine formulations at variousconcentrations Full Extrusions: ED (% LC) Full Extrusions: ED (mg) 32.0mg/mL 21.3 mg/mL 10.7 mg/mL 32.0 mg/mL 21.3 mg/mL 10.7 mg/mL NicotineNicotine Nicotine Nicotine Nicotine Nicotine ED # Bitartrate BitartrateBitartrate Bitartrate Bitartrate Bitartrate  1 56.2 55.6 59.5 0.90 0.590.32  2 59.4 59.7 62.6 0.95 0.64 0.34  3 56.9 61.8 60.4 0.91 0.66 0.32 4 56.6 60.4 62.9 0.91 0.64 0.34  5 57.7 60.9 65.3 0.92 0.65 0.35  658.1 64.8 62.3 0.93 0.69 0.33  7 63.6 57.9 60.0 1.02 0.62 0.32  8 60.365.2 58.1 0.96 0.69 0.31  9 54.6 59.9 66.1 0.87 0.64 0.35 10 58.4 58.067.9 0.93 0.62 0.36 11 59.8 60.1 66.8 0.96 0.64 0.36 12 63.8 63.0 61.81.02 0.67 0.33 13 60.0 59.7 64.6 0.96 0.64 0.35 14 63.1 62.8 65.0 1.010.67 0.35 15 61.2 63.7 56.0 0.98 0.68 0.30 16 64.5 66.0 62.0 1.03 0.700.33 17 62.7 62.0 65.2 1.00 0.66 0.35 18 65.5 65.7 63.7 1.05 0.70 0.3419 58.1 64.2 65.1 0.93 0.68 0.35 20 59.9 62.0 58.0 0.96 0.66 0.31 Mean60.0 61.7 62.7 0.96 0.66 0.34 SD 3.0 2.8 3.2 0.05 0.03 0.02 % RSD 5.14.6 5.1 5.1 4.6 5.1

Development of Dose-Titration Capabilities

Partial Extrusion of a Single AERx® Strip

Partial extrusion of an AERx® Strip was carried out by altering thesettings for the piston position, to program it to aerosolize only aportion of the contents of the AERx® Strip. Testing was done usingnicotine formulations, with the results being presented in Table 11. Thedelivered dose in percent of emitted dose at the three levels was 36.1,30.5 and 33.4 with standard deviations of 1.3, 1.1 and 1.3 respectively.This corresponds to a nicotine dose of 0.33 mg, 0.28 mg and 0.30 mg atthe three dose levels respectively.

Altering the Concentration of Nicotine in AERx® Strip

The emitted dose and particle size distribution of nicotine formulationsat various concentrations was evaluated. In order to keep the delivereddose constant, the range of concentrations tested were matched to theresults of the aerosol performance studies from partial extrusiondiscussed above. Results are presented in Table 13. The percent emitteddose for the successive concentrations of 32.0, 21.3 and 10.7 mg/mL was60.0, 61.7 and 62.7 with the standard deviations being 3.0, 2.8 and 3.2respectively. The corresponding delivered nicotine dose at the threeconcentrations was calculated to be 0.96 mg, 0.66 mg and 0.34 mg withstandard deviations of 0.05, 0.03 and 0.02 respectively.

TABLE 13 Emitted dose summary Emitted Drug Dose to the lung FormulationType of Extrusion (N = 20) % ED (SD) (mg) [FPF_(3.5) = 0.78] (mg) 32.0mg/mL Nicotine Partial Dose Level 1 20.4 (1.36) 0.33 0.26 Bitartrate, pH3.0 Partial Dose Level 2 37.9 (1.96) 0.61 0.48 Partial Dose Level 3 56.4(2.53) 0.90 0.71 10.7 mg/mL Nicotine Full Extrusion 62.7 (3.22) 0.340.27 Bitartrate, pH 3.0 21.3 mg/mL Nicotine Full Extrusion 61.7 (2.82)0.66 0.51 Bitartrate, pH 3.0 32.0 mg/mL Nicotine Full Extrusion 60.0(3.04) 0.96 0.75 Bitartrate, pH 3.0

Optimizing particle size distribution of the aerosol droplets ofnicotine formulations generated using AERx®

Particle size distribution (PSD) is a key determinant of the regionallung deposition of inhaled aerosols. A cascade impactor (Series 20-800Mark II, Thermo Andersen), which size selectively collects the aerosolby inertial impaction on a series of stages, was used to characterizethe aerosol PSD. The PSD was characterized in terms of Mass MedianAerodynamic Diameter (MMAD) and Geometric Standard Deviation (σg). MMADdenotes the particle size at which half of the total aerosol mass iscontained in larger particles and half in smaller particles. The σgindicates the variability of aerosol particle sizes. An aerosol composedof identical size particles would have a σg of 1.0; σg of ≦1.3 isconsidered monodisperse; σg of ≧1.3 is considered polydisperse.

We evaluated PSD with the optimized nicotine formulations. The MMADranged between 2.5-2.7 μm for the different combinations of device andformulation combinations (see Table 14). The GSD was 1.3, whichindicates the monodispersity of the aerosol. The influence of PSD onnicotine kinetics, efficiency, and success rates with the product wouldneed to be determined as part of the Phase II proposal. The fraction ofparticles under 3.5 μm is typically used to evaluate the fraction ofaerosol capable of deposition in the deep lung. The typical fineparticle fraction was about 80% (Table 14), indicating that the majorityof the deposited aerosol was capable of deep lung deposition, key to thesuccess of the therapy.

TABLE 14 Particle Size Distribution (PSD) summary Type of extrusionFormulation (N = 3) MMAD (SD) GSD (SD) FPF_(8.6) 32.0 mg/ml Nicotine 3partial 2.66 (0.04) 1.28 (0.01) 0.779 Bitartrate, pH 3.0 extrusions 32.0mg/ml Nicotine 1 partial 2.65 (0.03) 1.28 (0.01) 0.783 Bitartrate, pH3.0 extrusion 32.0 mg/ml Nicotine Full 2.49 (0.04) 1.35 (0.02) 0.762Bitartrate, pH 3.0 extrusions

Characterization of stability of nicotine formulations in AERx® Stripdosage forms

In the following set of experiments, the stability of the selectednicotine formulations was evaluated in AERx® Strip dosage forms.

Physical and chemical characterization of the selected formulations andaerosol performance upon storage in AERx® Strips for up to 1 month:

The primary storage condition for the strips was chosen to be 25° C./40%R.H., as the formulation selected was quite simple and did not requirerefrigerated storage. The strips were loaded with 50 μL of nicotineformulation, sealed and stored at 25° C./40% R.H., as well as at theaccelerated storage condition of 40° C./15% R.H. for up to 1 month. Theformulations in the strips were characterized for concentration, pH andcontent uniformity, in addition to measurement of aerosol performance(emitted dose, particle size distribution) with the AERx® Strips instorage. The results from the one month stability study indicatedmaintenance of pH, concentration, as well as aerosol performance overthe tested stability duration at the primary as well as acceleratedstorage condition (Tables 15 & 16). The emitted dose (ED) performance atboth the concentrations was within normal variability. The MMAD was 2.4and 2.8 to 2.9 for the two formulation strengths respectively; withGSD's of 1.3, indicating the monodispersity of the aerosol. The fineparticle fraction under 3.5 μm was about 82% for the 10.7 mg/mLformulation and 72% for the 32.0 mg/mL formulation. A high fraction ofthe emitted aerosol in the respirable range ensures that majority of theaerosol will result in deep lung deposition. The data indicatesacceptable stability of the formulations in AERx® strips for theduration of the stability study. In the next part of the developmentprogram, it will be important to finalize the final formulationconcentrations (dependent on the chosen dose reduction andcommercialization strategy) and generate stability data to support anyproposed clinical studies.

TABLE 15 Summary for Nicotine Bitartrate, 10.7 mg/ml in Aerx ® strips T= 4 weeks Test Attributes T = Initial 25° C./40% RH 40° C./15% RH pH (%RSD)  3.0 (0.2)  3.1 (0.0)  3.2 (0.4) Concentration, mg/mL (% RSD) 10.8(0.8) 10.7 (2.1) 10.7 (0.3) Content Uniformity Range, 97.0-100.0 N/A N/A% LC (% RSD) (1.2) Unit Dose, % LC (% RSD) 99.1 (1.2) 96.7 (1.7) 96.5(1.5) Emitted Dose, % LC (% RSD) 55.1 (4.9) 52.0 (5.8) 52.5 (3.0)Emitted Dose Uniformity, % 95.2-107.0 92.7-106.7 95.5-102.9 Mean ED (%RSD) (4.9) (5.8) (3.0) Particle Size Distribution 2.41, 1.31, 0.81, 44.62.42, 1.27, 0.85, 44.2 2.43, 1.28, 0.82, 43.1 [Record: MMAD (μm), GSD,FPF_(3.5), FPD (% LC)]

TABLE 16 Stability Summary for Nicotine Bitartrate, 32.0 mg/mL in AERx ®strips T = 4 weeks Test Attributes T = Initial 25° C./40% RH 40° C./15%RH pH (% RSD)  3.0 (0.0)  3.1 (0.5)  3.1 (0.8) Concentration, mg/mL (%RSD) 32.5 (0.9) 31.2 (1.0) 31.5 (1.0) Content Uniformity Range, % LC99.2-101.5 N/A N/A (% RSD) (0.8) Unit Dose, % LC (% RSD) 100.3 (0.8) 96.7 (0.7) 96.9 (0.5) Emitted Dose, % LC (% RSD) 58.3 (4.3) 52.8 (3.1)55.9 (1.3) Emitted Dose Uniformity, % Mean 94.0-106.8 95.6-103.197.9-101.5 ED (% RSD) (4.3) (3.1) (1.3) Particle Size Distribution[Record: 2.77, 1.27, 0.74, 43.1 2.87, 1.25, 0.70, 37.0 2.87, 1.25, 0.71,39.7 MMAD (÷m), GSD, FPF_(3.5), FPD (% LC)]

Conclusion

The example above supports the feasibility of delivery of nicotine forsmoking cessation using the AERx® System with an aqueous formulationthat was stable at room temperature for a period of at least a month(duration of stability study). The typical MMAD of the aerosols usingeither dose reduction strategy was 2.6 μm, whereas the GSD was 1.3. Thefine particle fraction was 80%, ensuring deposition of the majority ofthe emitted aerosol in the deep lung, mimicking smoking, and importantfor a successful smoking cessation product.

The preceding merely illustrates the principles of the invention. Itwill be appreciated that those skilled in the art will be able to devisevarious arrangements which, although not explicitly described or shownherein, embody the principles of the invention and are included withinits spirit and scope. Furthermore, all examples and conditional languagerecited herein are principally intended to aid the reader inunderstanding the principles of the invention and the conceptscontributed by the inventors to furthering the art, and are to beconstrued as being without limitation to such specifically recitedexamples and conditions. Moreover, all statements herein recitingprinciples, aspects, and embodiments of the invention as well asspecific examples thereof, are intended to encompass both structural andfunctional equivalents thereof. Additionally, it is intended that suchequivalents include both currently known equivalents and equivalentsdeveloped in the future, i.e., any elements developed that perform thesame function, regardless of structure. The scope of the presentinvention, therefore, is not intended to be limited to the exemplaryembodiments shown and described herein. Rather, the scope and spirit ofpresent invention is embodied by the appended claims.

1-20. (canceled)
 21. A tobacco-less kit, comprising: an aerosolizablenicotine formulation, comprising a carrier and nicotine in an amount toprovide a first nicotine arterial concentration in a mannersubstantially similar to a peak nicotine arterial plasma concentrationin a subject that is obtained by a cigarette; and a controlled releasenicotine formulation for delivery to maintain a second nicotine arterialconcentration in a manner substantially similar to a nicotine arterialplasma concentration in the subject that is maintained by the cigaretteafter the peak nicotine arterial plasma concentration.
 22. The kit ofclaim 21, wherein said first nicotine arterial concentration is at leastabout 10 ng/ml in the subject within 5 minutes of delivery of theaerosolizable formulation.
 23. The kit of claim 21, wherein said secondnicotine arterial concentration is at least about 5 ng/ml in the subjectfor at least 60 minutes after delivery of the controlled releaseformulation.
 24. The kit of claim 21, wherein delivering theaerosolizable formulation to said subject mimics an act of smoking. 25.The kit of claim 21, wherein aerosolizable formulation simulates thepharmacokinetics of nicotine delivered by a cigarette.
 26. The kit ofclaim 21, wherein the controlled release formulation is in a formselected from the group consisting of a Lozenge, gum, quick dissolvestrip, cream, gel, solid, transdermal patch, powder, liquid, suspension,and emulsion.
 27. The kit of claim 21, wherein the carrier is water. 28.The kit of claim 21, wherein the controlled release formulation is apiece of gum.
 29. The kit of claim 21, further comprising: an additionaldosage of a drug chosen from an antidepressant and an anxiolytic
 30. Thekit of claim 21, wherein the aerosolizable formulation is substantiallyfree base nicotine and wherein the formulation is in a form chosen froma solution, a powder, a suspension and an emulsion.
 31. The kit of claim21, wherein the aerosolizable formulation aerosolize into particleshaving an aerodynamic diameter of 0.5 to 12 microns.
 32. The kit ofclaim 29, wherein the average aerodynamic diameter is between 1 μm and 4μm.
 33. A kit of claim 21, further comprising: a plurality of containersof both aerosolized formulation and controlled release formulation. 34.The kit of claim 33, further comprising: an inhaler configured toaerosolize the aerosolizable formulation in the containers.
 35. The kitof claim 33, wherein said controlled release formulation is atransdermal patch.
 36. The kit of claim 33, wherein said controlledrelease formulation is a transmucosal formulation.
 37. The kit of claim33, further comprising: a plurality of doses of an antidepressant.